The uptake of naked or purified DNA by CELLS, usually meaning the process as it occurs in eukaryotic cells. It is analogous to bacterial transformation (TRANSFORMATION, BACTERIAL) and both are routinely employed in GENE TRANSFER TECHNIQUES.
Established cell cultures that have the potential to propagate indefinitely.
The sequence of PURINES and PYRIMIDINES in nucleic acids and polynucleotides. It is also called nucleotide sequence.
Extrachromosomal, usually CIRCULAR DNA molecules that are self-replicating and transferable from one organism to another. They are found in a variety of bacterial, archaeal, fungal, algal, and plant species. They are used in GENETIC ENGINEERING as CLONING VECTORS.
DNA sequences which are recognized (directly or indirectly) and bound by a DNA-dependent RNA polymerase during the initiation of transcription. Highly conserved sequences within the promoter include the Pribnow box in bacteria and the TATA BOX in eukaryotes.
Descriptions of specific amino acid, carbohydrate, or nucleotide sequences which have appeared in the published literature and/or are deposited in and maintained by databanks such as GENBANK, European Molecular Biology Laboratory (EMBL), National Biomedical Research Foundation (NBRF), or other sequence repositories.
Strongly cationic polymer that binds to certain proteins; used as a marker in immunology, to precipitate and purify enzymes and lipids. Synonyms: aziridine polymer; Epamine; Epomine; ethylenimine polymer; Montrek; PEI; Polymin(e).
A deoxyribonucleotide polymer that is the primary genetic material of all cells. Eukaryotic and prokaryotic organisms normally contain DNA in a double-stranded state, yet several important biological processes transiently involve single-stranded regions. DNA, which consists of a polysugar-phosphate backbone possessing projections of purines (adenine and guanine) and pyrimidines (thymine and cytosine), forms a double helix that is held together by hydrogen bonds between these purines and pyrimidines (adenine to thymine and guanine to cytosine).
DNA molecules capable of autonomous replication within a host cell and into which other DNA sequences can be inserted and thus amplified. Many are derived from PLASMIDS; BACTERIOPHAGES; or VIRUSES. They are used for transporting foreign genes into recipient cells. Genetic vectors possess a functional replicator site and contain GENETIC MARKERS to facilitate their selective recognition.
Cells grown in vitro from neoplastic tissue. If they can be established as a TUMOR CELL LINE, they can be propagated in cell culture indefinitely.
The biosynthesis of RNA carried out on a template of DNA. The biosynthesis of DNA from an RNA template is called REVERSE TRANSCRIPTION.
Enzymes that oxidize certain LUMINESCENT AGENTS to emit light (PHYSICAL LUMINESCENCE). The luciferases from different organisms have evolved differently so have different structures and substrates.
RNA sequences that serve as templates for protein synthesis. Bacterial mRNAs are generally primary transcripts in that they do not require post-transcriptional processing. Eukaryotic mRNA is synthesized in the nucleus and must be exported to the cytoplasm for translation. Most eukaryotic mRNAs have a sequence of polyadenylic acid at the 3' end, referred to as the poly(A) tail. The function of this tail is not known for certain, but it may play a role in the export of mature mRNA from the nucleus as well as in helping stabilize some mRNA molecules by retarding their degradation in the cytoplasm.
The introduction of functional (usually cloned) GENES into cells. A variety of techniques and naturally occurring processes are used for the gene transfer such as cell hybridization, LIPOSOMES or microcell-mediated gene transfer, ELECTROPORATION, chromosome-mediated gene transfer, TRANSFECTION, and GENETIC TRANSDUCTION. Gene transfer may result in genetically transformed cells and individual organisms.
Cells propagated in vitro in special media conducive to their growth. Cultured cells are used to study developmental, morphologic, metabolic, physiologic, and genetic processes, among others.
Any of the processes by which nuclear, cytoplasmic, or intercellular factors influence the differential control (induction or repression) of gene action at the level of transcription or translation.
Genes whose expression is easily detectable and therefore used to study promoter activity at many positions in a target genome. In recombinant DNA technology, these genes may be attached to a promoter region of interest.
A technique in which electric pulses of intensity in kilovolts per centimeter and of microsecond-to-millisecond duration cause a temporary loss of the semipermeability of CELL MEMBRANES, thus leading to ion leakage, escape of metabolites, and increased uptake by cells of drugs, molecular probes, and DNA.
Small double-stranded, non-protein coding RNAs (21-31 nucleotides) involved in GENE SILENCING functions, especially RNA INTERFERENCE (RNAi). Endogenously, siRNAs are generated from dsRNAs (RNA, DOUBLE-STRANDED) by the same ribonuclease, Dicer, that generates miRNAs (MICRORNAS). The perfect match of the siRNAs' antisense strand to their target RNAs mediates RNAi by siRNA-guided RNA cleavage. siRNAs fall into different classes including trans-acting siRNA (tasiRNA), repeat-associated RNA (rasiRNA), small-scan RNA (scnRNA), and Piwi protein-interacting RNA (piRNA) and have different specific gene silencing functions.
A cell line derived from cultured tumor cells.
Techniques and strategies which include the use of coding sequences and other conventional or radical means to transform or modify cells for the purpose of treating or reversing disease conditions.
Artificial, single or multilaminar vesicles (made from lecithins or other lipids) that are used for the delivery of a variety of biological molecules or molecular complexes to cells, for example, drug delivery and gene transfer. They are also used to study membranes and membrane proteins.
The phenotypic manifestation of a gene or genes by the processes of GENETIC TRANSCRIPTION and GENETIC TRANSLATION.
Proteins which bind to DNA. The family includes proteins which bind to both double- and single-stranded DNA and also includes specific DNA binding proteins in serum which can be used as markers for malignant diseases.
Endogenous substances, usually proteins, which are effective in the initiation, stimulation, or termination of the genetic transcription process.
The insertion of recombinant DNA molecules from prokaryotic and/or eukaryotic sources into a replicating vehicle, such as a plasmid or virus vector, and the introduction of the resultant hybrid molecules into recipient cells without altering the viability of those cells.
An enzyme that catalyzes the acetylation of chloramphenicol to yield chloramphenicol 3-acetate. Since chloramphenicol 3-acetate does not bind to bacterial ribosomes and is not an inhibitor of peptidyltransferase, the enzyme is responsible for the naturally occurring chloramphenicol resistance in bacteria. The enzyme, for which variants are known, is found in both gram-negative and gram-positive bacteria. EC
Single-stranded complementary DNA synthesized from an RNA template by the action of RNA-dependent DNA polymerase. cDNA (i.e., complementary DNA, not circular DNA, not C-DNA) is used in a variety of molecular cloning experiments as well as serving as a specific hybridization probe.
The first continuously cultured human malignant CELL LINE, derived from the cervical carcinoma of Henrietta Lacks. These cells are used for VIRUS CULTIVATION and antitumor drug screening assays.
Protein analogs and derivatives of the Aequorea victoria green fluorescent protein that emit light (FLUORESCENCE) when excited with ULTRAVIOLET RAYS. They are used in REPORTER GENES in doing GENETIC TECHNIQUES. Numerous mutants have been made to emit other colors or be sensitive to pH.
The order of amino acids as they occur in a polypeptide chain. This is referred to as the primary structure of proteins. It is of fundamental importance in determining PROTEIN CONFORMATION.
Positively charged atoms, radicals or groups of atoms which travel to the cathode or negative pole during electrolysis.
CELL LINES derived from the CV-1 cell line by transformation with a replication origin defective mutant of SV40 VIRUS, which codes for wild type large T antigen (ANTIGENS, POLYOMAVIRUS TRANSFORMING). They are used for transfection and cloning. (The CV-1 cell line was derived from the kidney of an adult male African green monkey (CERCOPITHECUS AETHIOPS).)
Processes that stimulate the GENETIC TRANSCRIPTION of a gene or set of genes.
A subfamily in the family MURIDAE, comprising the hamsters. Four of the more common genera are Cricetus, CRICETULUS; MESOCRICETUS; and PHODOPUS.
Recombinant proteins produced by the GENETIC TRANSLATION of fused genes formed by the combination of NUCLEIC ACID REGULATORY SEQUENCES of one or more genes with the protein coding sequences of one or more genes.
Identification of proteins or peptides that have been electrophoretically separated by blot transferring from the electrophoresis gel to strips of nitrocellulose paper, followed by labeling with antibody probes.
Any detectable and heritable change in the genetic material that causes a change in the GENOTYPE and which is transmitted to daughter cells and to succeeding generations.
Proteins prepared by recombinant DNA technology.
The intracellular transfer of information (biological activation/inhibition) through a signal pathway. In each signal transduction system, an activation/inhibition signal from a biologically active molecule (hormone, neurotransmitter) is mediated via the coupling of a receptor/enzyme to a second messenger system or to an ion channel. Signal transduction plays an important role in activating cellular functions, cell differentiation, and cell proliferation. Examples of signal transduction systems are the GAMMA-AMINOBUTYRIC ACID-postsynaptic receptor-calcium ion channel system, the receptor-mediated T-cell activation pathway, and the receptor-mediated activation of phospholipases. Those coupled to membrane depolarization or intracellular release of calcium include the receptor-mediated activation of cytotoxic functions in granulocytes and the synaptic potentiation of protein kinase activation. Some signal transduction pathways may be part of larger signal transduction pathways; for example, protein kinase activation is part of the platelet activation signal pathway.
A variation of the PCR technique in which cDNA is made from RNA via reverse transcription. The resultant cDNA is then amplified using standard PCR protocols.
The span of viability of a cell characterized by the capacity to perform certain functions such as metabolism, growth, reproduction, some form of responsiveness, and adaptability.
One of the mechanisms by which CELL DEATH occurs (compare with NECROSIS and AUTOPHAGOCYTOSIS). Apoptosis is the mechanism responsible for the physiological deletion of cells and appears to be intrinsically programmed. It is characterized by distinctive morphologic changes in the nucleus and cytoplasm, chromatin cleavage at regularly spaced sites, and the endonucleolytic cleavage of genomic DNA; (DNA FRAGMENTATION); at internucleosomal sites. This mode of cell death serves as a balance to mitosis in regulating the size of animal tissues and in mediating pathologic processes associated with tumor growth.
A peptide which is a homopolymer of lysine.
Short sequences (generally about 10 base pairs) of DNA that are complementary to sequences of messenger RNA and allow reverse transcriptases to start copying the adjacent sequences of mRNA. Primers are used extensively in genetic and molecular biology techniques.
High molecular weight insoluble polymers which contain functional anionic groups that are capable of undergoing exchange reactions with cations.
The parts of a macromolecule that directly participate in its specific combination with another molecule.
Derivatives of ammonium compounds, NH4+ Y-, in which all four of the hydrogens bonded to nitrogen have been replaced with hydrocarbyl groups. These are distinguished from IMINES which are RN=CR2.
A species of CERCOPITHECUS containing three subspecies: C. tantalus, C. pygerythrus, and C. sabeus. They are found in the forests and savannah of Africa. The African green monkey (C. pygerythrus) is the natural host of SIMIAN IMMUNODEFICIENCY VIRUS and is used in AIDS research.
A negative regulatory effect on physiological processes at the molecular, cellular, or systemic level. At the molecular level, the major regulatory sites include membrane receptors, genes (GENE EXPRESSION REGULATION), mRNAs (RNA, MESSENGER), and proteins.
Proteins found in the nucleus of a cell. Do not confuse with NUCLEOPROTEINS which are proteins conjugated with nucleic acids, that are not necessarily present in the nucleus.
CELL LINE derived from the ovary of the Chinese hamster, Cricetulus griseus (CRICETULUS). The species is a favorite for cytogenetic studies because of its small chromosome number. The cell line has provided model systems for the study of genetic alterations in cultured mammalian cells.
DNA that is complementary to the sense strand. (The sense strand has the same sequence as the mRNA transcript. The antisense strand is the template for mRNA synthesis.) Synthetic antisense DNAs are used to hybridize to complementary sequences in target RNAs or DNAs to effect the functioning of specific genes for investigative or therapeutic purposes.
Any of the processes by which nuclear, cytoplasmic, or intercellular factors influence the differential control of gene action in enzyme synthesis.
Cell lines whose original growing procedure consisted being transferred (T) every 3 days and plated at 300,000 cells per plate (J Cell Biol 17:299-313, 1963). Lines have been developed using several different strains of mice. Tissues are usually fibroblasts derived from mouse embryos but other types and sources have been developed as well. The 3T3 lines are valuable in vitro host systems for oncogenic virus transformation studies, since 3T3 cells possess a high sensitivity to CONTACT INHIBITION.
Deoxyribonucleic acid that makes up the genetic material of viruses.
The process in which substances, either endogenous or exogenous, bind to proteins, peptides, enzymes, protein precursors, or allied compounds. Specific protein-binding measures are often used as assays in diagnostic assessments.
Connective tissue cells which secrete an extracellular matrix rich in collagen and other macromolecules.
Diffusible gene products that act on homologous or heterologous molecules of viral or cellular DNA to regulate the expression of proteins.
The fission of a CELL. It includes CYTOKINESIS, when the CYTOPLASM of a cell is divided, and CELL NUCLEUS DIVISION.
A group of enzymes that catalyzes the hydrolysis of terminal, non-reducing beta-D-galactose residues in beta-galactosides. Deficiency of beta-Galactosidase A1 may cause GANGLIOSIDOSIS, GM1.
Within a eukaryotic cell, a membrane-limited body which contains chromosomes and one or more nucleoli (CELL NUCLEOLUS). The nuclear membrane consists of a double unit-type membrane which is perforated by a number of pores; the outermost membrane is continuous with the ENDOPLASMIC RETICULUM. A cell may contain more than one nucleus. (From Singleton & Sainsbury, Dictionary of Microbiology and Molecular Biology, 2d ed)
Derivatives of phosphatidic acids in which the phosphoric acid is bound in ester linkage to an ethanolamine moiety. Complete hydrolysis yields 1 mole of glycerol, phosphoric acid and ethanolamine and 2 moles of fatty acids.
Eukaryotic cell line obtained in a quiescent or stationary phase which undergoes conversion to a state of unregulated growth in culture, resembling an in vitro tumor. It occurs spontaneously or through interaction with viruses, oncogenes, radiation, or drugs/chemicals.
Any of the processes by which nuclear, cytoplasmic, or intercellular factors influence the differential control of gene action in neoplastic tissue.
Promoter-specific RNA polymerase II transcription factor that binds to the GC box, one of the upstream promoter elements, in mammalian cells. The binding of Sp1 is necessary for the initiation of transcription in the promoters of a variety of cellular and viral GENES.
A gene silencing phenomenon whereby specific dsRNAs (RNA, DOUBLE-STRANDED) trigger the degradation of homologous mRNA (RNA, MESSENGER). The specific dsRNAs are processed into SMALL INTERFERING RNA (siRNA) which serves as a guide for cleavage of the homologous mRNA in the RNA-INDUCED SILENCING COMPLEX. DNA METHYLATION may also be triggered during this process.
Techniques where DNA is delivered directly into organelles at high speed using projectiles coated with nucleic acid, shot from a helium-powered gun (gene gun). One of these techniques involves immunization by DNA VACCINES, which delivers DNA-coated gold beads to the epidermis.
Nucleic acid sequences involved in regulating the expression of genes.
Short fragments of DNA or RNA that are used to alter the function of target RNAs or DNAs to which they hybridize.
Cis-acting DNA sequences which can increase transcription of genes. Enhancers can usually function in either orientation and at various distances from a promoter.
Proteins which are involved in the phenomenon of light emission in living systems. Included are the "enzymatic" and "non-enzymatic" types of system with or without the presence of oxygen or co-factors.
The introduction of a phosphoryl group into a compound through the formation of an ester bond between the compound and a phosphorus moiety.
All of the processes involved in increasing CELL NUMBER including CELL DIVISION.
A positive regulatory effect on physiological processes at the molecular, cellular, or systemic level. At the molecular level, the major regulatory sites include membrane receptors, genes (GENE EXPRESSION REGULATION), mRNAs (RNA, MESSENGER), and proteins.
Genetically engineered MUTAGENESIS at a specific site in the DNA molecule that introduces a base substitution, or an insertion or deletion.
Detection of RNA that has been electrophoretically separated and immobilized by blotting on nitrocellulose or other type of paper or nylon membrane followed by hybridization with labeled NUCLEIC ACID PROBES.
The process of intracellular viral multiplication, consisting of the synthesis of PROTEINS; NUCLEIC ACIDS; and sometimes LIPIDS, and their assembly into a new infectious particle.
Relating to the size of solids.
Genes that are introduced into an organism using GENE TRANSFER TECHNIQUES.
Technique using an instrument system for making, processing, and displaying one or more measurements on individual cells obtained from a cell suspension. Cells are usually stained with one or more fluorescent dyes specific to cell components of interest, e.g., DNA, and fluorescence of each cell is measured as it rapidly transverses the excitation beam (laser or mercury arc lamp). Fluorescence provides a quantitative measure of various biochemical and biophysical properties of the cell, as well as a basis for cell sorting. Other measurable optical parameters include light absorption and light scattering, the latter being applicable to the measurement of cell size, shape, density, granularity, and stain uptake.
Products of proto-oncogenes. Normally they do not have oncogenic or transforming properties, but are involved in the regulation or differentiation of cell growth. They often have protein kinase activity.
A family of non-enveloped viruses infecting mammals (MASTADENOVIRUS) and birds (AVIADENOVIRUS) or both (ATADENOVIRUS). Infections may be asymptomatic or result in a variety of diseases.
Ubiquitous, inducible, nuclear transcriptional activator that binds to enhancer elements in many different cell types and is activated by pathogenic stimuli. The NF-kappa B complex is a heterodimer composed of two DNA-binding subunits: NF-kappa B1 and relA.
Nanometer-sized particles that are nanoscale in three dimensions. They include nanocrystaline materials; NANOCAPSULES; METAL NANOPARTICLES; DENDRIMERS, and QUANTUM DOTS. The uses of nanoparticles include DRUG DELIVERY SYSTEMS and cancer targeting and imaging.
Nucleotide sequences, usually upstream, which are recognized by specific regulatory transcription factors, thereby causing gene response to various regulatory agents. These elements may be found in both promoter and enhancer regions.
Fatty acids which are unsaturated in only one position.
Mutant mice homozygous for the recessive gene "nude" which fail to develop a thymus. They are useful in tumor studies and studies on immune responses.
Use of restriction endonucleases to analyze and generate a physical map of genomes, genes, or other segments of DNA.
In vitro method for producing large amounts of specific DNA or RNA fragments of defined length and sequence from small amounts of short oligonucleotide flanking sequences (primers). The essential steps include thermal denaturation of the double-stranded target molecules, annealing of the primers to their complementary sequences, and extension of the annealed primers by enzymatic synthesis with DNA polymerase. The reaction is efficient, specific, and extremely sensitive. Uses for the reaction include disease diagnosis, detection of difficult-to-isolate pathogens, mutation analysis, genetic testing, DNA sequencing, and analyzing evolutionary relationships.
Interruption or suppression of the expression of a gene at transcriptional or translational levels.
Cell changes manifested by escape from control mechanisms, increased growth potential, alterations in the cell surface, karyotypic abnormalities, morphological and biochemical deviations from the norm, and other attributes conferring the ability to invade, metastasize, and kill.
Deletion of sequences of nucleic acids from the genetic material of an individual.
Forms to which substances are incorporated to improve the delivery and the effectiveness of drugs. Drug carriers are used in drug-delivery systems such as the controlled-release technology to prolong in vivo drug actions, decrease drug metabolism, and reduce drug toxicity. Carriers are also used in designs to increase the effectiveness of drug delivery to the target sites of pharmacological actions. Liposomes, albumin microspheres, soluble synthetic polymers, DNA complexes, protein-drug conjugates, and carrier erythrocytes among others have been employed as biodegradable drug carriers.
Progressive restriction of the developmental potential and increasing specialization of function that leads to the formation of specialized cells, tissues, and organs.
Transport proteins that carry specific substances in the blood or across cell membranes.
Microscopy of specimens stained with fluorescent dye (usually fluorescein isothiocyanate) or of naturally fluorescent materials, which emit light when exposed to ultraviolet or blue light. Immunofluorescence microscopy utilizes antibodies that are labeled with fluorescent dye.
An electrophoretic technique for assaying the binding of one compound to another. Typically one compound is labeled to follow its mobility during electrophoresis. If the labeled compound is bound by the other compound, then the mobility of the labeled compound through the electrophoretic medium will be retarded.
Elements of limited time intervals, contributing to particular results or situations.
The functional hereditary units of VIRUSES.
Compounds formed by the joining of smaller, usually repeating, units linked by covalent bonds. These compounds often form large macromolecules (e.g., BIOPOLYMERS; PLASTICS).
Any of the processes by which cytoplasmic factors influence the differential control of gene action in viruses.
Biologically active DNA which has been formed by the in vitro joining of segments of DNA from different sources. It includes the recombination joint or edge of a heteroduplex region where two recombining DNA molecules are connected.
Proteins which are found in membranes including cellular and intracellular membranes. They consist of two types, peripheral and integral proteins. They include most membrane-associated enzymes, antigenic proteins, transport proteins, and drug, hormone, and lectin receptors.
Conversion of an inactive form of an enzyme to one possessing metabolic activity. It includes 1, activation by ions (activators); 2, activation by cofactors (coenzymes); and 3, conversion of an enzyme precursor (proenzyme or zymogen) to an active enzyme.
A group of deoxyribonucleotides (up to 12) in which the phosphate residues of each deoxyribonucleotide act as bridges in forming diester linkages between the deoxyribose moieties.
Small double-stranded, non-protein coding RNAs, 21-25 nucleotides in length generated from single-stranded microRNA gene transcripts by the same RIBONUCLEASE III, Dicer, that produces small interfering RNAs (RNA, SMALL INTERFERING). They become part of the RNA-INDUCED SILENCING COMPLEX and repress the translation (TRANSLATION, GENETIC) of target RNA by binding to homologous 3'UTR region as an imperfect match. The small temporal RNAs (stRNAs), let-7 and lin-4, from C. elegans, are the first 2 miRNAs discovered, and are from a class of miRNAs involved in developmental timing.
Genes whose gain-of-function alterations lead to NEOPLASTIC CELL TRANSFORMATION. They include, for example, genes for activators or stimulators of CELL PROLIFERATION such as growth factors, growth factor receptors, protein kinases, signal transducers, nuclear phosphoproteins, and transcription factors. A prefix of "v-" before oncogene symbols indicates oncogenes captured and transmitted by RETROVIRUSES; the prefix "c-" before the gene symbol of an oncogene indicates it is the cellular homolog (PROTO-ONCOGENES) of a v-oncogene.
A category of nucleic acid sequences that function as units of heredity and which code for the basic instructions for the development, reproduction, and maintenance of organisms.
A continuous cell line of high contact-inhibition established from NIH Swiss mouse embryo cultures. The cells are useful for DNA transfection and transformation studies. (From ATCC [Internet]. Virginia: American Type Culture Collection; c2002 [cited 2002 Sept 26]. Available from
A generic term for fats and lipoids, the alcohol-ether-soluble constituents of protoplasm, which are insoluble in water. They comprise the fats, fatty oils, essential oils, waxes, phospholipids, glycolipids, sulfolipids, aminolipids, chromolipids (lipochromes), and fatty acids. (Grant & Hackh's Chemical Dictionary, 5th ed)
Proteins found in any species of virus.
Cells that line the inner and outer surfaces of the body by forming cellular layers (EPITHELIUM) or masses. Epithelial cells lining the SKIN; the MOUTH; the NOSE; and the ANAL CANAL derive from ectoderm; those lining the RESPIRATORY SYSTEM and the DIGESTIVE SYSTEM derive from endoderm; others (CARDIOVASCULAR SYSTEM and LYMPHATIC SYSTEM) derive from mesoderm. Epithelial cells can be classified mainly by cell shape and function into squamous, glandular and transitional epithelial cells.
Proteins whose abnormal expression (gain or loss) are associated with the development, growth, or progression of NEOPLASMS. Some neoplasm proteins are tumor antigens (ANTIGENS, NEOPLASM), i.e. they induce an immune reaction to their tumor. Many neoplasm proteins have been characterized and are used as tumor markers (BIOMARKERS, TUMOR) when they are detectable in cells and body fluids as monitors for the presence or growth of tumors. Abnormal expression of ONCOGENE PROTEINS is involved in neoplastic transformation, whereas the loss of expression of TUMOR SUPPRESSOR PROTEINS is involved with the loss of growth control and progression of the neoplasm.
Family of retrovirus-associated DNA sequences (ras) originally isolated from Harvey (H-ras, Ha-ras, rasH) and Kirsten (K-ras, Ki-ras, rasK) murine sarcoma viruses. Ras genes are widely conserved among animal species and sequences corresponding to both H-ras and K-ras genes have been detected in human, avian, murine, and non-vertebrate genomes. The closely related N-ras gene has been detected in human neuroblastoma and sarcoma cell lines. All genes of the family have a similar exon-intron structure and each encodes a p21 protein.
A genus of the family Muridae consisting of eleven species. C. migratorius, the grey or Armenian hamster, and C. griseus, the Chinese hamster, are the two species used in biomedical research.
A light microscopic technique in which only a small spot is illuminated and observed at a time. An image is constructed through point-by-point scanning of the field in this manner. Light sources may be conventional or laser, and fluorescence or transmitted observations are possible.
Used as a support for ion-exchange chromatography.
Compounds or agents that combine with an enzyme in such a manner as to prevent the normal substrate-enzyme combination and the catalytic reaction.
A primary malignant neoplasm of epithelial liver cells. It ranges from a well-differentiated tumor with EPITHELIAL CELLS indistinguishable from normal HEPATOCYTES to a poorly differentiated neoplasm. The cells may be uniform or markedly pleomorphic, or form GIANT CELLS. Several classification schemes have been suggested.
Proteins which maintain the transcriptional quiescence of specific GENES or OPERONS. Classical repressor proteins are DNA-binding proteins that are normally bound to the OPERATOR REGION of an operon, or the ENHANCER SEQUENCES of a gene until a signal occurs that causes their release.
A subfield of acoustics dealing in the radio frequency range higher than acoustic SOUND waves (approximately above 20 kilohertz). Ultrasonic radiation is used therapeutically (DIATHERMY and ULTRASONIC THERAPY) to generate HEAT and to selectively destroy tissues. It is also used in diagnostics, for example, ULTRASONOGRAPHY; ECHOENCEPHALOGRAPHY; and ECHOCARDIOGRAPHY, to visually display echoes received from irradiated tissues.
The parts of a transcript of a split GENE remaining after the INTRONS are removed. They are spliced together to become a MESSENGER RNA or other functional RNA.
A cell line generated from human embryonic kidney cells that were transformed with human adenovirus type 5.
Change brought about to an organisms genetic composition by unidirectional transfer (TRANSFECTION; TRANSDUCTION, GENETIC; CONJUGATION, GENETIC, etc.) and incorporation of foreign DNA into prokaryotic or eukaryotic cells by recombination of part or all of that DNA into the cell's genome.
The sequential correspondence of nucleotides in one nucleic acid molecule with those of another nucleic acid molecule. Sequence homology is an indication of the genetic relatedness of different organisms and gene function.
Histochemical localization of immunoreactive substances using labeled antibodies as reagents.
The movement of cells from one location to another. Distinguish from CYTOKINESIS which is the process of dividing the CYTOPLASM of a cell.
The outward appearance of the individual. It is the product of interactions between genes, and between the GENOTYPE and the environment.
The rate dynamics in chemical or physical systems.
Proteins that are coded by immediate-early genes, in the absence of de novo protein synthesis. The term was originally used exclusively for viral regulatory proteins that were synthesized just after viral integration into the host cell. It is also used to describe cellular proteins which are synthesized immediately after the resting cell is stimulated by extracellular signals.
A group of enzymes that catalyzes the phosphorylation of serine or threonine residues in proteins, with ATP or other nucleotides as phosphate donors.
Ribonucleic acid that makes up the genetic material of viruses.
A species of POLYOMAVIRUS originally isolated from Rhesus monkey kidney tissue. It produces malignancy in human and newborn hamster kidney cell cultures.
Deacetylated CHITIN, a linear polysaccharide of deacetylated beta-1,4-D-glucosamine. It is used in HYDROGEL and to treat WOUNDS.
Adherence of cells to surfaces or to other cells.
A strain of albino rat used widely for experimental purposes because of its calmness and ease of handling. It was developed by the Sprague-Dawley Animal Company.
A multiprotein complex composed of the products of c-jun and c-fos proto-oncogenes. These proteins must dimerize in order to bind to the AP-1 recognition site, also known as the TPA-responsive element (TRE). AP-1 controls both basal and inducible transcription of several genes.
The biosynthesis of PEPTIDES and PROTEINS on RIBOSOMES, directed by MESSENGER RNA, via TRANSFER RNA that is charged with standard proteinogenic AMINO ACIDS.
A method (first developed by E.M. Southern) for detection of DNA that has been electrophoretically separated and immobilized by blotting on nitrocellulose or other type of paper or nylon membrane followed by hybridization with labeled NUCLEIC ACID PROBES.
Short fragments of DNA that are used to alter the function of target RNAs or DNAs to which they hybridize.
The degree of similarity between sequences of amino acids. This information is useful for the analyzing genetic relatedness of proteins and species.
Test for tissue antigen using either a direct method, by conjugation of antibody with fluorescent dye (FLUORESCENT ANTIBODY TECHNIQUE, DIRECT) or an indirect method, by formation of antigen-antibody complex which is then labeled with fluorescein-conjugated anti-immunoglobulin antibody (FLUORESCENT ANTIBODY TECHNIQUE, INDIRECT). The tissue is then examined by fluorescence microscopy.
A polynucleotide consisting essentially of chains with a repeating backbone of phosphate and ribose units to which nitrogenous bases are attached. RNA is unique among biological macromolecules in that it can encode genetic information, serve as an abundant structural component of cells, and also possesses catalytic activity. (Rieger et al., Glossary of Genetics: Classical and Molecular, 5th ed)
A cultured line of C3H mouse FIBROBLASTS that do not adhere to one another and do not express CADHERINS.
Glycoproteins found on the membrane or surface of cells.
An inheritable change in cells manifested by changes in cell division and growth and alterations in cell surface properties. It is induced by infection with a transforming virus.
The complex series of phenomena, occurring between the end of one CELL DIVISION and the end of the next, by which cellular material is duplicated and then divided between two daughter cells. The cell cycle includes INTERPHASE, which includes G0 PHASE; G1 PHASE; S PHASE; and G2 PHASE, and CELL DIVISION PHASE.
An enzyme capable of hydrolyzing highly polymerized DNA by splitting phosphodiester linkages, preferentially adjacent to a pyrimidine nucleotide. This catalyzes endonucleolytic cleavage of DNA yielding 5'-phosphodi- and oligonucleotide end-products. The enzyme has a preference for double-stranded DNA.
Polymers of ETHYLENE OXIDE and water, and their ethers. They vary in consistency from liquid to solid depending on the molecular weight indicated by a number following the name. They are used as SURFACTANTS, dispersing agents, solvents, ointment and suppository bases, vehicles, and tablet excipients. Some specific groups are NONOXYNOLS, OCTOXYNOLS, and POLOXAMERS.
Immunologic method used for detecting or quantifying immunoreactive substances. The substance is identified by first immobilizing it by blotting onto a membrane and then tagging it with labeled antibodies.
The relationship between the dose of an administered drug and the response of the organism to the drug.
Small encapsulated gas bubbles (diameters of micrometers) that can be used as CONTRAST MEDIA, and in other diagnostic and therapeutic applications. Upon exposure to sufficiently intense ultrasound, microbubbles will cavitate, rupture, disappear, release gas content. Such characteristics of the microbubbles can be used to enhance diagnostic tests, dissolve blood clots, and deliver drugs or genes for therapy.
A large lobed glandular organ in the abdomen of vertebrates that is responsible for detoxification, metabolism, synthesis and storage of various substances.
RNA molecules which hybridize to complementary sequences in either RNA or DNA altering the function of the latter. Endogenous antisense RNAs function as regulators of gene expression by a variety of mechanisms. Synthetic antisense RNAs are used to effect the functioning of specific genes for investigative or therapeutic purposes.
Linear POLYPEPTIDES that are synthesized on RIBOSOMES and may be further modified, crosslinked, cleaved, or assembled into complex proteins with several subunits. The specific sequence of AMINO ACIDS determines the shape the polypeptide will take, during PROTEIN FOLDING, and the function of the protein.
The sequence at the 3' end of messenger RNA that does not code for product. This region contains transcription and translation regulating sequences.
Serologic tests in which a positive reaction manifested by visible CHEMICAL PRECIPITATION occurs when a soluble ANTIGEN reacts with its precipitins, i.e., ANTIBODIES that can form a precipitate.
Sequences of DNA in the genes that are located between the EXONS. They are transcribed along with the exons but are removed from the primary gene transcript by RNA SPLICING to leave mature RNA. Some introns code for separate genes.
The part of a cell that contains the CYTOSOL and small structures excluding the CELL NUCLEUS; MITOCHONDRIA; and large VACUOLES. (Glick, Glossary of Biochemistry and Molecular Biology, 1990)
Viruses whose hosts are bacterial cells.
The artificial induction of GENE SILENCING by the use of RNA INTERFERENCE to reduce the expression of a specific gene. It includes the use of DOUBLE-STRANDED RNA, such as SMALL INTERFERING RNA and RNA containing HAIRPIN LOOP SEQUENCE, and ANTI-SENSE OLIGONUCLEOTIDES.
Tumors or cancer of the LIVER.
Process of generating a genetic MUTATION. It may occur spontaneously or be induced by MUTAGENS.
Polymers made up of a few (2-20) nucleotides. In molecular genetics, they refer to a short sequence synthesized to match a region where a mutation is known to occur, and then used as a probe (OLIGONUCLEOTIDE PROBES). (Dorland, 28th ed)
Different forms of a protein that may be produced from different GENES, or from the same gene by ALTERNATIVE SPLICING.
A superfamily of PROTEIN-SERINE-THREONINE KINASES that are activated by diverse stimuli via protein kinase cascades. They are the final components of the cascades, activated by phosphorylation by MITOGEN-ACTIVATED PROTEIN KINASE KINASES, which in turn are activated by mitogen-activated protein kinase kinase kinases (MAP KINASE KINASE KINASES).
Experimental transplantation of neoplasms in laboratory animals for research purposes.
The region of DNA which borders the 5' end of a transcription unit and where a variety of regulatory sequences are located.
The level of protein structure in which combinations of secondary protein structures (alpha helices, beta sheets, loop regions, and motifs) pack together to form folded shapes called domains. Disulfide bridges between cysteines in two different parts of the polypeptide chain along with other interactions between the chains play a role in the formation and stabilization of tertiary structure. Small proteins usually consist of only one domain but larger proteins may contain a number of domains connected by segments of polypeptide chain which lack regular secondary structure.
Body organ that filters blood for the secretion of URINE and that regulates ion concentrations.
An enzyme that catalyzes the conversion of ATP and thymidine to ADP and thymidine 5'-phosphate. Deoxyuridine can also act as an acceptor and dGTP as a donor. (From Enzyme Nomenclature, 1992) EC
A mutation caused by the substitution of one nucleotide for another. This results in the DNA molecule having a change in a single base pair.
Proteins encoded by homeobox genes (GENES, HOMEOBOX) that exhibit structural similarity to certain prokaryotic and eukaryotic DNA-binding proteins. Homeodomain proteins are involved in the control of gene expression during morphogenesis and development (GENE EXPRESSION REGULATION, DEVELOPMENTAL).
Domesticated bovine animals of the genus Bos, usually kept on a farm or ranch and used for the production of meat or dairy products or for heavy labor.
A protein-serine-threonine kinase that is activated by PHOSPHORYLATION in response to GROWTH FACTORS or INSULIN. It plays a major role in cell metabolism, growth, and survival as a core component of SIGNAL TRANSDUCTION. Three isoforms have been described in mammalian cells.
Nuclear phosphoprotein encoded by the p53 gene (GENES, P53) whose normal function is to control CELL PROLIFERATION and APOPTOSIS. A mutant or absent p53 protein has been found in LEUKEMIA; OSTEOSARCOMA; LUNG CANCER; and COLORECTAL CANCER.
A method for determining the sequence specificity of DNA-binding proteins. DNA footprinting utilizes a DNA damaging agent (either a chemical reagent or a nuclease) which cleaves DNA at every base pair. DNA cleavage is inhibited where the ligand binds to DNA. (from Rieger et al., Glossary of Genetics: Classical and Molecular, 5th ed)
The complete genetic complement contained in a DNA or RNA molecule in a virus.
A CELL LINE derived from human T-CELL LEUKEMIA and used to determine the mechanism of differential susceptibility to anti-cancer drugs and radiation.
Structurally related forms of an enzyme. Each isoenzyme has the same mechanism and classification, but differs in its chemical, physical, or immunological characteristics.
The process of moving proteins from one cellular compartment (including extracellular) to another by various sorting and transport mechanisms such as gated transport, protein translocation, and vesicular transport.
A process whereby multiple RNA transcripts are generated from a single gene. Alternative splicing involves the splicing together of other possible sets of EXONS during the processing of some, but not all, transcripts of the gene. Thus a particular exon may be connected to any one of several alternative exons to form a mature RNA. The alternative forms of mature MESSENGER RNA produce PROTEIN ISOFORMS in which one part of the isoforms is common while the other parts are different.
Genes that influence the PHENOTYPE both in the homozygous and the heterozygous state.
A theoretical representative nucleotide or amino acid sequence in which each nucleotide or amino acid is the one which occurs most frequently at that site in the different sequences which occur in nature. The phrase also refers to an actual sequence which approximates the theoretical consensus. A known CONSERVED SEQUENCE set is represented by a consensus sequence. Commonly observed supersecondary protein structures (AMINO ACID MOTIFS) are often formed by conserved sequences.
Sequences of DNA or RNA that occur in multiple copies. There are several types: INTERSPERSED REPETITIVE SEQUENCES are copies of transposable elements (DNA TRANSPOSABLE ELEMENTS or RETROELEMENTS) dispersed throughout the genome. TERMINAL REPEAT SEQUENCES flank both ends of another sequence, for example, the long terminal repeats (LTRs) on RETROVIRUSES. Variations may be direct repeats, those occurring in the same direction, or inverted repeats, those opposite to each other in direction. TANDEM REPEAT SEQUENCES are copies which lie adjacent to each other, direct or inverted (INVERTED REPEAT SEQUENCES).
A group of simple proteins that yield basic amino acids on hydrolysis and that occur combined with nucleic acid in the sperm of fish. Protamines contain very few kinds of amino acids. Protamine sulfate combines with heparin to form a stable inactive complex; it is used to neutralize the anticoagulant action of heparin in the treatment of heparin overdose. (From Merck Index, 11th ed; Martindale, The Extra Pharmacopoeia, 30th ed, p692)
The lipid- and protein-containing, selectively permeable membrane that surrounds the cytoplasm in prokaryotic and eukaryotic cells.
Family of RNA viruses that infects birds and mammals and encodes the enzyme reverse transcriptase. The family contains seven genera: DELTARETROVIRUS; LENTIVIRUS; RETROVIRUSES TYPE B, MAMMALIAN; ALPHARETROVIRUS; GAMMARETROVIRUS; RETROVIRUSES TYPE D; and SPUMAVIRUS. A key feature of retrovirus biology is the synthesis of a DNA copy of the genome which is integrated into cellular DNA. After integration it is sometimes not expressed but maintained in a latent state (PROVIRUSES).
Intracellular receptors that can be found in the cytoplasm or in the nucleus. They bind to extracellular signaling molecules that migrate through or are transported across the CELL MEMBRANE. Many members of this class of receptors occur in the cytoplasm and are transported to the CELL NUCLEUS upon ligand-binding where they signal via DNA-binding and transcription regulation. Also included in this category are receptors found on INTRACELLULAR MEMBRANES that act via mechanisms similar to CELL SURFACE RECEPTORS.
Compound such as LUMINESCENT PROTEINS that cause or emit light (PHYSICAL LUMINESCENCE).
Characteristic restricted to a particular organ of the body, such as a cell type, metabolic response or expression of a particular protein or antigen.
The species Oryctolagus cuniculus, in the family Leporidae, order LAGOMORPHA. Rabbits are born in burrows, furless, and with eyes and ears closed. In contrast with HARES, rabbits have 22 chromosome pairs.
Cellular DNA-binding proteins encoded by the c-jun genes (GENES, JUN). They are involved in growth-related transcriptional control. There appear to be three distinct functions: dimerization (with c-fos), DNA-binding, and transcriptional activation. Oncogenic transformation can take place by constitutive expression of c-jun.
A human liver tumor cell line used to study a variety of liver-specific metabolic functions.
A class of proteins that were originally identified by their ability to bind the DNA sequence CCAAT. The typical CCAAT-enhancer binding protein forms dimers and consists of an activation domain, a DNA-binding basic region, and a leucine-rich dimerization domain (LEUCINE ZIPPERS). CCAAT-BINDING FACTOR is structurally distinct type of CCAAT-enhancer binding protein consisting of a trimer of three different subunits.
Biochemical identification of mutational changes in a nucleotide sequence.
Protein kinases that catalyze the PHOSPHORYLATION of TYROSINE residues in proteins with ATP or other nucleotides as phosphate donors.

11q23.1 and 11q25-qter YACs suppress tumour growth in vivo. (1/63574)

Frequent allelic deletion at chromosome 11q22-q23.1 has been described in breast cancer and a number of other malignancies, suggesting putative tumour suppressor gene(s) within the approximately 8 Mb deleted region. In addition, we recently described another locus, at the 11q25-qter region, frequently deleted in breast cancer, suggesting additional tumour suppressor gene(s) in this approximately 2 Mb deleted region. An 11q YAC contig was accessed and three YACs, one containing the candidate gene ATM at 11q23.1, and two contiguous YACs (overlapping for approximately 400-600 kb) overlying most of the 11q25 deleted region, were retrofitted with a G418 resistance marker and transfected into murine A9 fibrosarcoma cells. Selected A9 transfectant clones (and control untransfected and 'irrelevant' alphoid YAC transfectant A9 clones) were assayed for in vivo tumorigenicity in athymic female Balb c-nu/nu mice. All the 11q YAC transfectant clones demonstrated significant tumour suppression compared to the control untransfected and 'irrelevant' YAC transfected A9 cells. These results define two discrete tumour suppressor loci on chromosome 11q by functional complementation, one to a approximately 1.2 Mb region on 11q23.1 (containing the ATM locus) and another to a approximately 400-600 kb subterminal region on 11q25-qter.  (+info)

TIF1gamma, a novel member of the transcriptional intermediary factor 1 family. (2/63574)

We report the cloning and characterization of a novel member of the Transcriptional Intermediary Factor 1 (TIF1) gene family, human TIF1gamma. Similar to TIF1alpha and TIF1beta, the structure of TIF1beta is characterized by multiple domains: RING finger, B boxes, Coiled coil, PHD/TTC, and bromodomain. Although structurally related to TIF1alpha and TIF1beta, TIF1gamma presents several functional differences. In contrast to TIF1alpha, but like TIF1beta, TIF1 does not interact with nuclear receptors in yeast two-hybrid or GST pull-down assays and does not interfere with retinoic acid response in transfected mammalian cells. Whereas TIF1alpha and TIF1beta were previously found to interact with the KRAB silencing domain of KOX1 and with the HP1alpha, MODI (HP1beta) and MOD2 (HP1gamma) heterochromatinic proteins, suggesting that they may participate in a complex involved in heterochromatin-induced gene repression, TIF1gamma does not interact with either the KRAB domain of KOX1 or the HP1 proteins. Nevertheless, TIF1gamma, like TIF1alpha and TIF1beta, exhibits a strong silencing activity when tethered to a promoter. Since deletion of a novel motif unique to the three TIF1 proteins, called TIF1 signature sequence (TSS), abrogates transcriptional repression by TIF1gamma, this motif likely participates in TIF1 dependent repression.  (+info)

Expression of the naturally occurring truncated trkB neurotrophin receptor induces outgrowth of filopodia and processes in neuroblastoma cells. (3/63574)

We have investigated the effects of the truncated trkB receptor isoform T1 (trkB.T1) by transient transfection into mouse N2a neuroblastoma cells. We observed that expression of trkB.T1 leads to a striking change in cell morphology characterized by outgrowth of filopodia and processes. A similar morphological response was also observed in SH-SY5Y human neuroblastoma cells and NIH3T3 fibroblasts transfected with trkB.T1. N2a cells lack endogenous expression of trkB isoforms, but express barely detectable amounts of its ligands, brain-derived neurotrophic factor (BDNF) and neurotrophin-4 (NT-4). The morphological change was ligand-independent, since addition of exogenous BDNF or NT-4 or blockade of endogenous trkB ligands did not influence this response. Filopodia and process outgrowth was significantly suppressed when full-length trkB.TK+ was cotransfected together with trkB.T1 and this inhibitory effect was blocked by tyrosine kinase inhibitor K252a. Transfection of trkB.T1 deletion mutants showed that the morphological response is dependent on the extracellular, but not the intracellular domain of the receptor. Our results suggest a novel ligand-independent role for truncated trkB in the regulation of cellular morphology.  (+info)

B-MYB transactivates its own promoter through SP1-binding sites. (4/63574)

B-MYB is an ubiquitous protein required for mammalian cell growth. In this report we show that B-MYB transactivates its own promoter through a 120 bp segment proximal to the transcription start site. The B-MYB-responsive element does not contain myb-binding sites and gel-shift analysis shows that SP1, but not B-MYB, protein contained in SAOS2 cell extracts binds to the 120 bp B-myb promoter fragment. B-MYB-dependent transactivation is cooperatively increased in the presence of SP1, but not SP3 overexpression. When the SP1 elements of the B-myb promoter are transferred in front of a heterologous promoter, an increased response to B-MYB results. In contrast, c-MYB, the prototype member of the Myb family, is not able to activate the luciferase construct containing the SP1 elements. With the use of an SP1-GAL4 fusion protein, we have determined that the cooperative activation occurs through the domain A of SP1. These observations suggest that B-MYB functions as a coactivator of SP1, and that diverse combinations of myb and SP1 sites may dictate the responsiveness of myb-target genes to the various members of the myb family.  (+info)

Arrestin function in G protein-coupled receptor endocytosis requires phosphoinositide binding. (5/63574)

Internalization of agonist-activated G protein-coupled receptors is mediated by non-visual arrestins, which also bind to clathrin and are therefore thought to act as adaptors in the endocytosis process. Phosphoinositides have been implicated in the regulation of intracellular receptor trafficking, and are known to bind to other coat components including AP-2, AP180 and COPI coatomer. Given these observations, we explored the possibility that phosphoinositides play a role in arrestin's function as an adaptor. High-affinity binding sites for phosphoinositides in beta-arrestin (arrestin2) and arrestin3 (beta-arrestin2) were identified, and dissimilar effects of phosphoinositide and inositol phosphate on arrestin interactions with clathrin and receptor were characterized. Alteration of three basic residues in arrestin3 abolished phosphoinositide binding with complete retention of clathrin and receptor binding. Unlike native protein, upon agonist activation, this mutant arrestin3 expressed in COS1 cells neither supported beta2-adrenergic receptor internalization nor did it concentrate in coated pits, although it was recruited to the plasma membrane. These findings indicate that phosphoinositide binding plays a critical regulatory role in delivery of the receptor-arrestin complex to coated pits, perhaps by providing, with activated receptor, a multi-point attachment of arrestin to the plasma membrane.  (+info)

The MAP kinase ERK2 inhibits the cyclic AMP-specific phosphodiesterase HSPDE4D3 by phosphorylating it at Ser579. (6/63574)

The extracellular receptor stimulated kinase ERK2 (p42(MAPK))-phosphorylated human cAMP-specific phosphodiesterase PDE4D3 at Ser579 and profoundly reduced ( approximately 75%) its activity. These effects could be reversed by the action of protein phosphatase PP1. The inhibitory state of PDE4D3, engendered by ERK2 phosphorylation, was mimicked by the Ser579-->Asp mutant form of PDE4D3. In COS1 cells transfected to express PDE4D3, challenge with epidermal growth factor (EGF) caused the phosphorylation and inhibition of PDE4D3. This effect was blocked by the MEK inhibitor PD98059 and was not apparent using the Ser579-->Ala mutant form of PDE4D3. Challenge of HEK293 and F442A cells with EGF led to the PD98059-ablatable inhibition of endogenous PDE4D3 and PDE4D5 activities. EGF challenge of COS1 cells transfected to express PDE4D3 increased cAMP levels through a process ablated by PD98059. The activity of the Ser579-->Asp mutant form of PDE4D3 was increased by PKA phosphorylation. The transient form of the EGF-induced inhibition of PDE4D3 is thus suggested to be due to feedback regulation by PKA causing the ablation of the ERK2-induced inhibition of PDE4D3. We identify a novel means of cross-talk between the cAMP and ERK signalling pathways whereby cell stimuli that lead to ERK2 activation may modulate cAMP signalling.  (+info)

Coupling of the cell cycle and myogenesis through the cyclin D1-dependent interaction of MyoD with cdk4. (7/63574)

Proliferating myoblasts express the muscle determination factor, MyoD, throughout the cell cycle in the absence of differentiation. Here we show that a mitogen-sensitive mechanism, involving the direct interaction between MyoD and cdk4, restricts myoblast differentiation to cells that have entered into the G0 phase of the cell cycle under mitogen withdrawal. Interaction between MyoD and cdk4 disrupts MyoD DNA-binding, muscle-specific gene activation and myogenic conversion of 10T1/2 cells independently of cyclin D1 and the CAK activation of cdk4. Forced induction of cyclin D1 in myotubes results in the cytoplasmic to nuclear translocation of cdk4. The specific MyoD-cdk4 interaction in dividing myoblasts, coupled with the cyclin D1-dependent nuclear targeting of cdk4, suggests a mitogen-sensitive mechanism whereby cyclin D1 can regulate MyoD function and the onset of myogenesis by controlling the cellular location of cdk4 rather than the phosphorylation status of MyoD.  (+info)

Assembly requirements of PU.1-Pip (IRF-4) activator complexes: inhibiting function in vivo using fused dimers. (8/63574)

Gene expression in higher eukaryotes appears to be regulated by specific combinations of transcription factors binding to regulatory sequences. The Ets factor PU.1 and the IRF protein Pip (IRF-4) represent a pair of interacting transcription factors implicated in regulating B cell-specific gene expression. Pip is recruited to its binding site on DNA by phosphorylated PU.1. PU.1-Pip interaction is shown to be template directed and involves two distinct protein-protein interaction surfaces: (i) the ets and IRF DNA-binding domains; and (ii) the phosphorylated PEST region of PU.1 and a lysine-requiring putative alpha-helix in Pip. Thus, a coordinated set of protein-protein and protein-DNA contacts are essential for PU.1-Pip ternary complex assembly. To analyze the function of these factors in vivo, we engineered chimeric repressors containing the ets and IRF DNA-binding domains connected by a flexible POU domain linker. When stably expressed, the wild-type fused dimer strongly repressed the expression of a rearranged immunoglobulin lambda gene, thereby establishing the functional importance of PU.1-Pip complexes in B cell gene expression. Comparative analysis of the wild-type dimer with a series of mutant dimers distinguished a gene regulated by PU.1 and Pip from one regulated by PU.1 alone. This strategy should prove generally useful in analyzing the function of interacting transcription factors in vivo, and for identifying novel genes regulated by such complexes.  (+info)

Cell Line,cell type,cell line specific transfection reagents,EZ Biosystems is a worldwide provider of transfection and gene expression product and services. Simpler, Faster, and Easier are EZ Biosystems promises to scientists in the life science research community. One of the major focuses of EZ Biosystems has been the research, development, and manufacture of next generation transfection reagents ---- Avalanche Transfection Reagent series by applying combinatorial chemistry, molecular biology, and cell biology expertise. This series includes: Avalanche-Cell Type/Cell Line-specific Transfection Reagent series: The pre-optimized transfection Reagent series that ensures the highest transfection efficiency and viability for over 100 cell types/cell lines. Avalanche-Omni Transfection Reagent: Broadest spectrum in vitro gene and siRNA transfection with exceptional transfection efficiency and viability. Avalanche-in vivo Transfection Reagent: Extremely powerful for gene functional studies and RNA
SignaGen Laboratories, A Gene Delivery Company Providing Custom AAV Adenovirus Lentivirus Production Services & Manufacturing DNA/siRNA Transfection Reagents... : > Pre-optimized Transfection Reagents - > In Vitro DNA Transfection Reagents > In Vivo Transfection Reagents > Pre-optimized Transfection Reagents > In Vitro siRNA Transfection Reagents > Transfection Accessories > Pre-made Adenovirus > shRNA/miRNA AAV Service > Adenovirus Production Service > shRNA/miRNA Adenovirus Svc > AAV Production Service > Ready-to-package Adenovirus > Pre-made AAVs > Pre-made Lentivirus > Lentivirus Production Service SignaGen Laboratories, DNA/siRNA Transfection Reagents, Custom Adenovirus Production Service, Custom AAV Production Service, Custom Lentivirus Production Service
Lipofectamine® RNAiMAX Transfection Reagent provides the highest transfection efficiencies on the widest variety of cell types for siRNA-mediated gene knockdown experiments. Lipofectamine® RNAiMAX is a proprietary RNAi-specific cationic lipid formulation designed specifically for the delivery of siRNA and miRNA into all cell types.With Lipofectamine® RNAiMAX Transfection Reagent you will get: • Superior transfection efficiency requiring lower RNAi concentrations, leading to more effective gene knockdown with minimal nonspecific effects• Easy optimization due to minimal cytotoxicity across a 10-fold concentration range of transfection reagent• Superior transfection efficiencies for miRNA antagonists and mimics• Compatibility with a broad range of cell types, providing the most versatile approach to all of your gene silencing experiments• A simple and rapid protocol for consistent and reproducible resultsHigh knockdown in a wide range of cellsLipofectamine® RNAiMAX Transfection Reagent
SignaGen Laboratories, A Gene Delivery Company Providing Custom AAV Adenovirus Lentivirus Production Services & Manufacturing DNA/siRNA Transfection Reagents... GenJet In Vitro DNA Transfection Reagent for Rin Related Cells [SL100489-RIN] - Description GenJet DNA In Vitro Transfection Reagent for Rin is pre-optimized znd conditioned for transfecting Rin and related cells including Rin-14B, Rin-m5F, Rin-m and Rin-5F. Refer to the following optimal transfection conditions for maximal transfection efficiency on Rin related cells. GenJet reagent, 1.0 ml, is sufficient for 300 to
Altogen Biosystems provides in vivo transfection reagents, over 100 pre-optimized in vitro transfection kits for cell lines and primary cells, and electroporation delivery products. Transfection reagents are highly efficient for DNA and siRNA transfection in vivo and in vitro. Altogen CRO offers in vivo RNAi services, tumor xenograft models, toxicology testing, stable cell line generation, and cell banking cryopreservation services
The choice of a transfection reagent often depends more upon the particular cell line than the substance being delivered into the cells. We have available four different lipid-based transfection reagents that have been specially formulated for delivery of siRNAs into cells. These DharmaFECT reagents are very mild yet have very high transfection rates into a wide variety of cell lines. You can find a list of cells weve tested with DharmaFECT here: http: //dharmacon. horizondiscovery. com/transfection/dharmafect-cell-type-guide/Another recommendation is to use a reagent that has been previously used in the cells of interest. Further optimization for siRNA delivery may be necessary. If no established protocol for the cells is available, a PubMed or HighWire search may help identify if a published protocol for the specific cell line or a similar cell line has been reported. In all instances, we recommend following the protocol provided by the manufacturer of the transfection reagent ...
Low transfection efficiency and low cell viability are the most frequent causes of unsuccessful gene silencing experiments. Through careful optimization--e.g. choosing the right transfection agent and transfection method--high levels of transfection efficiency can be achieved ,Optimizing,siRNA,Transfection,for,RNAi,biological,advanced biology technology,biology laboratory technology,biology device technology,latest biology technology
TY - JOUR. T1 - Protective immunity induced by B7.1+IL-12 transfected tumor cells is abrogated by tumor growth in an immune privileged site. AU - Chen, P. W.. AU - Uno, T.. AU - Geer, D. G.. AU - Podack, E. R.. AU - Ksander, B. R.. PY - 1996/12/1. Y1 - 1996/12/1. N2 - Mice immunized with B7.1+IL-12 transfected tumor cells possess systemic anti-tumor immunity that protects mice from a second subcutaneous tumor challenge of untransfected cells. This study determines if these immunized mice are protected from a second tumor challenge delivered to an immunologically privileged site. Our previous results indicated that tumors growing within the immunologically privileged anterior chamber of the eye (AC) of naive mice induce systemic T cell tolerance. In these experiments we determined if tolerance could be imposed after mice were previously sensitized. DBA/2 mice were given a subcutaneous immunization with 1 × 106 P815 B7.1+IL-12 cells. One month later, mice received a second tumor challenge of ...
Altogen Biosystems provides in vivo transfection reagents, over 100 pre-optimized in vitro transfection kits for cell lines and primary cells, and electroporation delivery products. Transfection reagents are highly efficient for DNA and siRNA transfection in vivo and in vitro. Altogen CRO offers in vivo RNAi services, tumor xenograft models, toxicology testing, stable cell line generation, and cell banking cryopreservation services
Images: Ret receptor knockdown using small interference RNA (siRNA) in podocytes. (A) Transfection efficiency: mouse podocytes were transfected with 100 nM concentrations of Ret siRNA or vehicle alone. (a) When podocytes were exposed to i-Fect alone, there was no toxicity. (a and b) A transfection efficiency of nearly 100% was achieved with 100 nM concentration of Ret siRNA (b). Co-transfection with a fluorescently tagged control siRNA was used to determine the transfection efficiency, and fluorescence microscopy revealed a perinuclear localization of the tagged RNA (b, arrowhead). (B) Western blot analysis of Ret after transfection: Ret immunoblotting (top) of WCL 2, 3, or 4 d after transfection revealed that Ret was downregulated within 2 d after transfection with 100 nM Ret siRNA. Transfection of control siRNA at day 4 served as a negative control, and the maximal knockdown of Ret was observed 4 d after transfection. GAPDH immunoblotting confirmed equal protein loading (bottom). ...
Roots Analysis has announced the addition of Non-Viral Transfection Reagents and Systems Market, 2020-2030 report to its list of offerings.. Over time, innovative approaches surrounding the development of potential therapies using non-viral transfection systems have prompted several companies to commercialize proprietary technologies to facilitate gene transfer into cells, via a variety of physical, chemical and other non-viral methods. The growing demand for safe and effective genetically engineered ATMPs is likely to further propel the opportunity for non-viral transfection system developers.. To order this 220+ page report, which features 100+ figures and 125+ tables, please visit the website. Key Market Insights. More than 110 companies claim to offer different types of non-viral transfection systems. The majority of players engaged in the development and commercialization of non-viral transfection systems offer reagents (52%), followed by companies offering electroporation-based ...
G-FECT™ DNA Transfection reagents are a series of powerful DNA transfection reagents with high efficiency DNA delivery and low cell toxicity into mammalian cells. G-FECT™ can be used for plasmid transfection, genome editing, virus production and more.
Forward transfection is a widely used method that works well for adherent cell types; however, if one is using suspension cells or a high-throughput format, reverse transfection, where cells are added to the plated transfection reagent complex, is a more appropriate approach. This method can be used to increase the throughput and reproducibility of a CRISPR-Cas9 screen by optimizing gene editing efficiency as demonstrated in our recently published application note: Optimization of reverse transfection of Dharmacon Edit-R synthetic crRNA and tracrRNA components with DharmaFECT transfection reagent in a Cas9-expressing cell line.. An un-cleavable ubiquitin moiety fused to EGFP allows constitutive degradation of the EGFP protein, while disruption of the proteasome components by functional protein knockout leads to accumulation of EGFP and detectable fluorescence. This functional knockout data shows that determining optimal transfection conditions prior to arrayed screening leads to high gene ...
siPORT NeoFX Transfection Agent 1 ml from Ambion,Gene silencing experiments often call for critical, but time-consuming optimization experiments. siPORT NeoFX Transfection Agent refines siRNA transfection protocols resulting in less optimization. siPORT NeoFXs lipid-based formulation can be used to efficiently transfect adherent cells as they are,biological,biology supply,biology supplies,biology product
Best transfection method/reagent for HeLa cells? - posted in Cell Biology: Im planning to do some transient transfection of HeLa cells. Aim is to do some CoIP and localization studies. I asked different fellows, some say Fugene is the best, some proposed TFX while other said, this is all crap, use Lipofectamine. Now im a little bit puzzled, which one i should buy... Which transfection method / reagent would you recommend?
Objectives and Background Insulin secretion entirely depends upon Ca2+ influx and sequestration into endoplasmic reticulum (ER) of In diabetes, SERCA2b is decreased within the gene transfected AMSCs for the pancreas of induced diabetes type 1 in rat. injected as with group III. Organizations I, II, IV and III were sacrified 3 weeks following verification of diabetes. Serological, histological, morphometric research and quantitative polymerase string reaction (qPCR) had been performed. Nuclear, cytoplasmic degenerative and intensive fibrotic changes had been detected within the islets of group II that regressed in organizations III and IV. Isolated islet calcium mineral, blood glucose, plasma insulin and qPCR had been confirmative. Conclusions AMSCs and gene transfected AMSCs therapy proved definite therapeutic effect, more obvious in response to gene transfected AMSCs. gene transfected AMSCs on the pancreas of streptozotocin (STZ)-induced diabetes type 1 in adult male albino rat and comparing it ...
Lipofectamine 2000 Transfection Reagent is a versatile transfection reagent that has been shown to effectively transfect the widest variety of adherent and suspension cell lines. Researchers use Lipofectamine 2000 Reagent for siRNA- and shRNA-based gene knockdown experiments, as well as for gene exp
Lipofectamine 2000 Transfection Reagent is a versatile transfection reagent that has been shown to effectively transfect the widest variety of adherent and suspension cell lines. Researchers use Lipofectamine 2000 Reagent for siRNA- and shRNA-based gene knockdown experiments, as well as for gene exp
GeneJuice® Transfection Reagent Non-lipid based chemical transfection reagent optimized for maximum transfection efficiency, ease-of-use, and minimal cytotoxicity on a wide variety of mammallian cells. - Find MSDS or SDS, a COA, data sheets and more information.
Lipo2000 Transfection Reagent Lipo 2000 Transfection Reagent is a proprietary formulation for transfecting nucleic acids into a wide range of eukaryotic cells. DNA lipofectamine 2000 complexes must be made in serum-free medium.
Ive done this, its no problem at all! (cells Ive done this on: U87 already transfected with 2 extra (selected) proteins and Hela cells already transfected with 2 proteins and 2 other sequences, thus a total of 4 different pieces of separate DNA integrated stably ...
TransIT®-Keratinocyte from Mirus Bio is a high efficiency, low toxicity, plasmid DNA transfection reagent, specifically optimised for keratinocyte transfection. Ideal for cells including NIKS (near-diploid immortalised keratinocytes) and primary keratinocytes, TransIT-Keratinocyte is suitable for both stable and transient transfections.. Benefits of using TransIT-Keratinocyte ...
Yes, antibiotics can be used during transfection with any of the Life Technologies lipid based reagents. However, the critical steps of the protocol need to be performed with the recommended OptiMEM Media and not culture media that contains any serum or antibiotics. The transfection protocol involves a two tube protocol and It is important to add the lipid reagent to OptiMEM in one tube and the DNA or RNA to OptiMEM in another tube and mix each tube thoroughly. Then, equal amounts from each tube should be mixed to form the transfection complex and incubated for five minutes at room temperature. The lipid-DNA or lipid-RNA complex can then be added directly to cells in complete culture media. Analysis of transfection results can be performed 24-48 hours post transfection. ...
InvivoGen provides LyoVec, a lyophilized cationic lipid-based transfection reagent made of phosphonolipid DTCPTA coupled with DiPPE, a neutral lipid that helps destabilizing membrane bilayers, therefore increasing the in vitro transfection efficiency of LyoVec.
The global transfection reagent & equipment market is expected to reach USD 1,086.02 million by 2022, according to a new report by Grand View Research, Inc. This anticipated growth in demand can be attributed to growing need for protein production, biopharmaceutical development, and vaccine research and development; all of which rely heavily on cytological R&D and transfection.. Complete Report Available @ Larger market entities are also involved in efforts to expand their market presence in the Asia Pacific region and tap the high potential for growth available. Expected growth of genomic and proteomic research is also a strong factor which will positively impact the growth in demand for transfection.. Further key findings from the study suggest:. ...
* This reagent has two solutions: Solution A - 0.5 ml, Solution B - 1.0 ml The TransPass™ R2 Transfection Reagent is a two component non-liposomal reagent combination developed specifically for efficient transfection of siRNA in a variety of mammalian cell lines that include “difficult to transfect cells such as primary cells
The goal of stable, long-term transfection is to isolate and propagate individual clones containing transfected DNA that has integrated into the cellular genome. Distinguishing nontransfected cells from those that have taken up exogenous DNA involves selective screening. This screening can be accomplished by drug selection when an appropriate drug-resistance marker is included in the transfected DNA. Alternatively, morphological transformation can be used as a selectable trait in certain cases. For example, bovine papilloma virus vectors produce a morphological change in transfected mouse CI127 cells.. Before using a particular drug for selection purposes, you will need to determine the amount of drug necessary to kill untransfected cells. This may vary greatly among cell types. Consult Current Protocols in Molecular Biology for additional information about designing experiments to test various drug concentrations and determine the amount needed to select resistant clones (i.e., generate a kill ...
줄기세포를 이용한 치료법은 광범위한 세포, 조직, 심지어 장기의 치료에까지 적용될 수 있다. 지방유래 줄기 세포는 다량 추출이 가능하다는 점과 치료 적용시 자가지방을 이용하게 됨으로써 면역반응을 줄일 수 있고, 다양한 세포로 분화가 가능하다. 하지만 충분한 양을 획득하기 위해 생체 밖에서 계대배양을 해야하는데, 이를 지속할 경우 줄기세포의 노화로 인해 줄기세포능을 잃는 어려움이 있다. 이러한 어려움을 개선시키기 위한 방안으로, 세포노화에 관여하는 것으로 알려진 Akt를 저해하는 PTEN 유전자를 선정하였다. PTEN 유전자를 지방 유래 성체줄기세포에 유전자치료는 지속적인 단백질 방출이 가능하기 때문에 한 번의 투여로 지속적인 치료효과를 얻을 수 있다는 장점이 있다. 줄기세포의 노화를 방지하면서 이러한 효과를 지속적으로 유지시켜줄 ...
Sigmas Universal Transfection Reagent is a unique formulation of a proprietary polymer blend used for transient and stable transfection of nucleic acids into various eukaryotic cell lines and hard-to-transfect primary cells. Fast and easy protocol is compatible with serum, serum-free medium and antibiotics
ATCC® Transfection reagents may be used for the transfer of nucleic acids for a variety of applications, including protein expression and the overexpression of mutant genes. Our transfection reagents are cationic lipid-based; due to their positive charge, these transfection reagents interact with the negatively charged backbone of nucleic acids and cell membranes. This effect, coupled with the hydrophobic nature of the associated lipids, allows the cationic lipid-DNA complex to be transported into eukaryotic cells. ATCC offers:. ...
Our i-FectTM Transfection Kit is used to study Epigenetics and pain. Heres yet another example: M. Leinders, b, N. Üçeyler, R.A. Pritchard, C. Sommer, L.S. Sorkin. Increased miR-132-3p expression is associated with chronic neuropathic pain. Experimental Neurology. Volume 283, Part A, September 2016, Pages 276-286...The inhibitor and mimetic were administered to awake rats via the it catheters. Prior to injection, active or mismatch inhibitors were mixed with (1:5 w/v) i-Fect™ in vivo transfection reagent(Neuromics, Edina, USA) to final doses of 5, 2 and 1 μg in 10 μl ...
Looking for a powerful and versatile DNA and siRNA transfection reagent? Try jetPRIME to obtain efficient and reliable scientific results! Request your jetPRIME sample immediately!
GeneX Plus Transfection Reagent is a high-performance, animal-origin free, broad spectrum reagent that provides exceptional transfection of plasmid DNA into mammalian cells.
Photoporation is a rapidly expanding technique for the introduction of macromolecules into single cells. However, there remains no study into the true efficiency of this procedure. Here, we present a detailed analysis of transfection efficiency and cell viability for femtosecond optical transfection using a titanium sapphire laser at 800 nm. Photoporation of 4000 Chinese Hamster ovary cells was performed, representing the largest optical transfection study reported to date. We have investigated a range of laser fluences at the cell membrane and, at 1.2 μJ/cm2, have found an average transfection efficiency of 50 ± 10%. Contrary to recent literature, in which 100% efficiency is claimed, our measure of efficiency accounts for all irradiated cells, including those lost as a result of laser treatment, thereby providing a true biological measure of the technique.. ©2006 Optical Society of America. Full Article , PDF Article ...
Transfection is the process of inserting genetic material, such as DNA and double stranded RNA, into mammalian cells. The insertion of DNA into a cell...
METAFECTENE EASY + Fast, easy and effective transfection reagent for mammalian cells For ordering information, MSDS, publications and application notes see Description Cat. No. Size METAFECTENE
Dear flow cytometry people, A problem with the viability of sorted cells was detected in our lab. We have been asked to sort a fibroblast cell line, IOT 1/2. Cells have been transfected using fugene, and GFP expressing cells (about 15%)and non-expressing cells were sorted into 15-ml falcon tubes filled with medium (5 ml) and maintained in ice. Cells before sorting had a clear FS/SS pattern, and viability using PI was about 95-98%. The instrument used was a MoFlo; laser is a Coherent Enterprise operated at 150 mW at 488nm. In the first attempts, a 90-micron tip at 20 PSI was used. A commercial PBS from DakoCytomation was used as sheath fluid. The problem was detected with sorted cells: both positive and negative tubes showed few cells with a low FS signal when reanalysed, and viability using PI was lower than 15-20%. Then we changed to a 200-micron tip at 5 PSI, but number and viability of sorted cells remained very low. Non-transfected cells showed the same behaviour when sorted. I wonder if it ...
Supplementary Materialscancers-12-01563-s001. MHC-I-negative murine tumor cell genes and lines from the IFN- transduction sign pathway are participating. Fhit-transfected tumor cells demonstrated immunogenic extremely, being rejected with a T lymphocyte-mediated immune system response. Strikingly, this immune Vitamin A system rejection was even more regular in females than in men. The immune system response generated secured hosts against the tumor development of non-transfected cells and against various other tumor cells inside our murine tumor PEPCK-C model. Finally, we also noticed a Vitamin A direct relationship between FHIT appearance and HLA-I surface area expression in individual breasts tumors. Recovery of Fhit appearance on MHC course I harmful tumor cells could be a good immunotherapeutic strategy and could even become an individualized immunotherapeutic vaccine. 0.05. A two-tailed Learners 0.05. A two-tailed Learners 0.001, Fisher check) (Body 3A; Body S7A). Small male ...
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Transfection Reagent and Equipment Market Trend, Development Analysis 2020 - Top Countries Data Analysis by Industry Share, Future Prospects, Industry Scope, Opportunity, Boosting Strategies, COVID-19 Impact by ...
Transfection Reagents at Biomol ► Life Science Shop for Research Scientists and Procurement Managers ► For Universities, Research Institutes, and Biotech Companies.
AMSBIO offers transfection reagents including GeneSilencer® with a unique cationic lipid formulation for efficient transfection of siRNA into mammalian cells.
Optimize transfection efficiencies for Cell Line Development; simple cell counting and viability measurements; and total cell counts for beverage contamination quality control.
FectoPRO®: Improving transient CHO and HEK-293 Expression systems with a powerful transfection solution for high protein production yields.. The development process for producing a biotherapeutic protein usually begins with generating a high-performing stable cell line which can be used for manufacturing. As this step takes a lot of time, transient transfection offers a great alternative to quickly produce milligram to gram quantities of recombinant proteins and antibodies. A various number culture medium are available for performing transient protein production in both CHO and HEK cells but the limiting factor is often the transfection reagent. Thats why we have developed a novel technologically advanced transfection solution called FectoPRO®. Here we show that FectoPRO® outperforms currently available PEI-based and lipid-based transfection reagents in all the transient expression systems tested, offering great transfection efficiency and amazing protein yields.. ...
To date, methods for large-scale transient gene expression (TGE) in cultivated mammalian cells have focused on two transfection vehicles: polyethylenimine (PEI) and calcium phosphate (CaPi). Both have been shown to result in high transfection efficiencies at scales beyond 10 L. Unfortunately, both approaches yield higher levels of recombinant protein (r-protein) in the presence of serum than in its absence. Since serum is a major cost factor and an obstacle to protein purification, our goal was to develop a large-scale TGE process for Chinese hamster ovary (CHO) cells in the absence of serum. CHO-DG44 cells were cultivated and transfected in a chemically defined medium using linear 25 kDa PEI as a transfection vehicle. Parameters that were optimized included the DNA amount, the DNA-to-PEI ratio, the timing and solution conditions for complex formation, the transfection medium, and the cell density at the time of transfection. The highest levels of r-protein expression were observed when cultures ...
TY - JOUR. T1 - Production of HIV virus-like particles by transient transfection of CAP-T cells at bioreactor scale avoiding medium replacement. AU - Gutiérrez-Granados, Sonia. AU - Farràs, Queralt. AU - Hein, Kerstin. AU - Fuenmayor, Javier. AU - Félez, Pablo. AU - Segura, Mercedes. AU - Gòdia, Francesc. PY - 2017/12/10. Y1 - 2017/12/10. N2 - © 2017 Elsevier B.V. Human-derived CAP-T cell line has been demonstrated to be a powerful platform for high-titer production of HIV virus-like particles (VLPs) by PEI-mediated transient transfection. Scale-up of transfection processes is key to ensure the necessary quantities for pre-clinical and clinical testing. One of the major operational challenges of large-scale transient transfection is the medium replacement step that is often required before transfection. In this work, CAP-T cells were cultured in 1 L bioreactor with addition of sodium bicarbonate and surface aeration, which were observed to improve cell state for transfection. Remarkably, ...
In this paper, Beckman Coulter Life Sciences how can the transfection efficiency be improved by using the Biomek i7 Automated Workstation.
The TGEX™-eGFP vector is a green fluorescent reporter designed to monitor transfection efficiency during transient gene expression in mammalian cell suspensions culture. TGEX™-eGFP vector can be used as a single component during transfection to analyze transfection efficiency or at a lower level, between 5% and 10% of the total DNA, to measure efficiency during transient gene expression.. ...
(EMAILWIRE.COM, May 22, 2017 ) Browse 134 market data tables and 43 figures spread through 209 pages and in-depth TOC on Transfection Reagents and Equipment Market Early buyers will receive 10%...
ExoFectin® Plasmid DNA-into-Exosome Kit (DNA transfection kit for exosome) is a unique blend of polymers designed for the delivery of plasmid DNA into exosomes. This transfection reagent provides highly efficient transfection of plasmid DNA into exosomes, allowing modified exosomes to carry and deliver plasmid DNA into target cells. This kit provides the following advantages:. ...
TY - JOUR. T1 - PEGylation improves the receptor-mediated transfection efficiency of peptide-targeted, self-assembling, anionic nanocomplexes. AU - Tagalakis, Aristides. AU - Kenny, G. AU - Bienemann, A.S.. AU - McCarthy, D. AU - Munye, M.M.. AU - Taylor, H. AU - Wyatt, M.J.. AU - Lyhtgoe, M.F.. AU - White, E.A.. AU - Hart, S.L.. PY - 2014/1/28. Y1 - 2014/1/28. N2 - Non-viral vector formulations comprise typically complexes of nucleic acids with cationic polymers or lipids. However, for in vivo applications cationic formulations suffer from problems of poor tissue penetration, non-specific binding to cells, interaction with serum proteins and cell adhesion molecules and can lead to inflammatory responses. Anionic formulations may provide a solution to these problems but they have not been developed to the same extent as cationic formulations due to difficulties of nucleic acid packaging and poor transfection efficiency. We have developed novel PEGylated, anionic nanocomplexes containing cationic ...
ViaFect Transfection Reagent is a high-performing, low-toxicity reagent designed to efficiently transfect DNA into a wide variety of cell lines.
In 2015, the reagents segment is expected to account for the largest share of the transfection reagents and equipment market, by product; the biochemical method segment is expected to account for the largest share of the global transfection reagents and equipment market, by method; while the biomedical research segment is expected to account for the largest share of the transfection reagents and equipment market, by application.. In 2015, North America is expected to account for the largest share of the global transfection reagents and equipment market, followed by Europe, Asia-Pacific, and rest of the world (RoW). In future, the transfection reagents and equipment market is expected to witness the highest growth rate in the Asia-Pacific region, with emphasis on India, China, and Japan. High growth in these countries can be attributed to the increase in research activities conducted in these regions, rapid expansion of the biotechnology and pharmaceutical industry, government as well as private ...
MACSfectin™ Reagent is applicable for plasmid DNA, mRNA, and siRNA transfection. For enrichment of transfected cells, the MACSelect™ System, which is based on the renowned MACS® Technology, helps to tackle low transfection efficiencies. It is uses the transiently expressed truncated human CD4, the truncated mouse MHC class I H-2Kk, and truncated human LNGFR as surface markers to select transfected cells. Transduction of cells with only low-titer preparations can be achieved by using MACSductin™ Reagent. It consists of polycationic, magnetic beads that help to efficiently transduce primary cells and cell lines using adeno- or retro-/lentiviral vectors ...
Vacic download tristes P technologies are for Differentiating random-coil people. acid of risk into dirged proposals may affect transduced by solitary proteins, doubt research, DEAE-dextran or acetylation. Promega comes download theories spoken on several siblings( TransFast™ Transfection Reagent and ViaFect™ Transfection Reagent), good lessons( FuGENE® 6 invention Reagent and FuGENE® HD Transfection Reagent) and accordance interaction( ProFection® Mammalian Transfection System).
The role of PTEN in cell spreading was also examined in transiently transfected primary human fibroblasts and DBTRG-05MG cells, a glioblastoma cell line with a 204-base pair deletion inPTEN (1). Cells expressing the various constructs were detected by using green fluorescent protein (GFP) as a marker (10). Again, PTEN inhibited cell spreading on FN (Fig. 2, B and C). In human fibroblasts, the effect was maximal at 20 to 30 min, with only 22 ± 8% of PTEN-overexpressing cells spread at 30 min compared with 60 ± 8% of nontransfected cells or cells with control GFP-plasmid (P , 0.001 to 0.005) (Fig. 2C). The majority of cells had spread by 2 hours, however, indicating that PTEN overexpression delayed but did not prevent spreading. Nevertheless, the surface area covered byPTEN-expressing cells remained less than that of controls (see below). In DBTRG-05MG cells, exogenous expression ofPTEN also delayed spreading; even after 18 hours, only 64% of the cells had spread, and the extent of spreading of ...
* found in: Convoy™ Transfection Reagent, Penetratin 1 Peptide, GeneTran™ III Tranfection Reagent, ConvoyTM is new generation cationic polymer gene..
WE ARE A DISTRIBUTING PARTNER OF ScreenFect - InCella ScreenFect® transfection reagents can efficiently transfect a variety of cells with minimal cytoxicit...
Fast SnapFect cell transfection with superior viability and efficiency New way to transfect cells with RNA, DNA, CRISPR Rapid SnapFect Transfection in Flow. (A) The bio-orthogonal mediated nucleic acid transfection strategy is fast and can be performed in a microfluidic format. The mixing of the keto tailored cells wit
Plasmid transfection in bovine cells: Optimization using a realtime monitoring of green fluorescent protein and effect on gene reporter assay.
Middle East and Africa Transfection Reagents and Equipment market size was around USD 28.62 million in 2016. It is expected to grow at a CAGR of 5.6 % to reach USD 37.61 million by 2021. It has the lowest market share of 4%.
Im interested in transient transfection of primary mouse embryo fibroblasts. Does anyone have any experience with this and what are the best methods/protocols for high transfection efficiency (,40%)? Rizwan ...
Plasmid DNA Transfection - Information on DNA transfection including plasmid transfection, transient transfection and stable transfection. Includes protocols and articles related to DNA transfection. ...
Transfection is the introduction of nucleic acid molecules into cultured mammalian cells. Chemical transfection reagents provide tools for researchers in a variety of gene research applications.
The MaxCyte® STX™ Scalable Transfection System uses a proprietary flow electroporation technology that can transfect up to 1E10 cells with target, reporter and protein expression plasmids, as well as other molecules, in less than 30 minutes. Transfected cells can be assayed immediately or cryopreserved for future use. Here we demonstrate the use of the MaxCyte STX system in coordination with Cornings ® HYPERFlask® Cell Culture Vessel to provide an efficient and economical solution for culturing large numbers of adherent CHO cells before and after transfection. The HYPERFlask Cell Culture Vessel features Cornings HYPER (High Yield PERformance) technology, which utilizes a gas permeable film to provide gas exchange between the internal culture environment and the external atmospheric environment. The unique, space-saving, 10-layer film design results in 1720 cm2 cell growth surface area, which is approximately 10 times that of a normal T-175 flask. We also show that cells transfected with a ...
Materials. Recombinant neuregulin (rHRGβ1177-244, a peptide of HRGβ1 residues 177-244) was generously provided by Dr. M. Sliwkowski (Holmes et al., 1992). Anti-ERK kinase antibody was a gift from Dr. G. S. Feng. The mouse AChR ε-subunit cDNA was provided by Dr. R. Huganir. The c-fosand c-jun cDNAs were provided by Drs. T. Curran and A. Langley (Curran et al., 1987). Dr. R. Davis provided JNK1, MKK4, and p38 constructs (Whitmarsh et al., 1997), and Dr. M. Birrer provided the c-JUN mammalian expression constructs (Brown et al., 1994). PD98059, AG1478, and SB202190 were purchased from Calbiochem (La Jolla, CA). Rapamycin was from Research Biochemicals (Natick, MA). Cell culture media were purchased from Life Technologies (Gaithersburg, MD). Anti-FLAG antibody and all other chemicals were from Sigma (St. Louis, MO).. Cell culture and transfection procedures. Mouse muscle C2C12 cells were maintained as undifferentiated myoblasts in a nutrient-rich growth medium containing DMEM supplemented with ...
siRNA transfection is a powerful tool used to understand the mechanisms of gene regulation and molecular pathways. The following 10 tips will help you to optimize your siRNA transfection.
Previous studies have implicated Cx-mediated cell-to-cell coupling inβ -cell function (Bosco et al., 1995; Calabrese et al., 2001; Cao et al., 1997; Vozzi et al., 1995) and have suggested that adequate levels of specific Cx isoforms is required for the control of insulin secretion of permanent cell lines (Calabrese et al., 2001; Cao et al., 1997; Vozzi et al., 1995). As yet, however, it has not proved feasible to test this hypothesis directly in normal pancreatic β-cells, mostly because these highly differentiated cells proliferate poorly (Nielsen et al., 1989) and hence are not amenable to the stable expression of exogenous cDNAs by standard transfection procedures. Alternative approaches, which take advantage of viral vectors that can infect pancreatic islets, have been shown to be an effective solution to this problem (Curran et al., 2002; Gallichan et al., 1998; Leibowitz et al., 1999; Salmon et al., 2000). However, these approaches are still limited by the poor viability of β-cells ...
Previous studies have implicated Cx-mediated cell-to-cell coupling inβ -cell function (Bosco et al., 1995; Calabrese et al., 2001; Cao et al., 1997; Vozzi et al., 1995) and have suggested that adequate levels of specific Cx isoforms is required for the control of insulin secretion of permanent cell lines (Calabrese et al., 2001; Cao et al., 1997; Vozzi et al., 1995). As yet, however, it has not proved feasible to test this hypothesis directly in normal pancreatic β-cells, mostly because these highly differentiated cells proliferate poorly (Nielsen et al., 1989) and hence are not amenable to the stable expression of exogenous cDNAs by standard transfection procedures. Alternative approaches, which take advantage of viral vectors that can infect pancreatic islets, have been shown to be an effective solution to this problem (Curran et al., 2002; Gallichan et al., 1998; Leibowitz et al., 1999; Salmon et al., 2000). However, these approaches are still limited by the poor viability of β-cells ...
article{9e981584-ee0b-4f80-867b-a0b6814c78bd, author = {Belting, Mattias and Petersson, Per}, language = {eng}, pages = {281--286}, series = {Biochem. J.}, title = {Protective role for proteoglycans against cationic lipid cytotoxicity allowing optimal transfection efficiency in vitro}, year = {1999 ...
The optimization of the lab intern standard protocols was one of our fields on investigation. Besides optimizing the handling of our different cell lines and working steps, the transfection protocol was examined. One of the most remarkable lessons after several transfection performed was the crucial handling of the AAV293 cells. Once over approximately 80 % confluence the cells are no longer competent for transfection. Another achievement in method development was the determination of the optimal plasmid amounts. The best results were performed using 3.3 µg per plasmid and therefore this parameter was modified in our standard protocol. After transfecting the AAV293 we were able to detect the Ca2+-DNA conglomerates in the medium. The toxic side effects of these conglomerates were also confirmed. Not only the medium had to be changed, but also washing with PBS was essential to keep the cells alive. The most critical steps in transfection is the exact pH of the 2x Hepes buffered-saline (HBS) ...
Transfection[edit]. Bacterial spheroplasts, with suitable recombinant DNA inserted into it, can be used to transfect animal ...
"Polyplus transfection". CNRS. "Les quinze lauréats de la Médaille d'argent du CNRS 1998". Archived from the original ... These vectors are widely used as transfection agents for animal cells in culture, but also as drug-gene carriers in clinical ... To this end, he founded two biotechnology companies, Eurothéra (1994-97) and Polyplus-transfection (2001). Research & Sharing ...
Kingston, Robert E.; Chen, Claudia A.; Rose, John K. (2003). "Calcium Phosphate Transfection". Current Protocols in Molecular ... lipid-mediated DNA-transfection procedure". Proceedings of the National Academy of Sciences. 84 (21): 7413-7417. doi:10.1073/ ...
Potter, H (2003). "Transfection by Electroporation". Current Protocols in Molecular Biology. Arena CB, Sano MB, Rossmeisl JH, ... Electroporation has proven efficient for use on tissues in vivo, for in utero applications as well as in ovo transfection. ... Particularly, the electroporation allows for a more efficient transfection of DNA, RNA, shRNA, and all nucleic acids into the ... The process of introducing foreign DNA into eukaryotic cells is known as transfection. Electroporation is highly effective for ...
... for he also developed two methods of calcium phosphate transfection for transient and stable transfections. These two methods ... Kingston, Robert E.; Chen, Claudia A.; Rose, John K. (2003). "Calcium Phosphate Transfection". Current Protocols in Molecular ... Robert Kingston's advancements within the field of biotechnology extend into developing eukaryotic cell transfection protocols ...
Fanelli A (2016). "Transfection: In Vitro Transfection". Retrieved 5 December 2017. Jensen K, Anderson JA, Glass EJ (April 2014 ... The most common commercial reagents for transfection of siRNA are Lipofectamine and Neon Transfection. However it is not ... Gene knockdown by transfection of exogenous siRNA is often unsatisfactory because the effect is only transient, especially in ... siRNAs transfection into cells typically lowers the expression of many genes, however, the upregulation of genes is also ...
"Polybrene Infection / Transfection Reagent , TR-1003-G". Retrieved 2016-12-02. Hunkapiller, M. W.; Hood, ...
Gulick, Tod (2003). "Transfection Using DEAE-Dextran". Current Protocols in Cell Biology. 19: 20.4.1-20.4.10. doi:10.1002/ ... It is added to solution containing DNA meant for transfection. It binds and interacts with negatively charged DNA molecules and ... This procedure is highly suited for transient transfection used for various molecular biology studies. DEAE-Sepharose, DEAE-650 ...
Membrane activity and transfection ability of amphipathic polycations as a function of alkyl group size. Bioconjug Chem. 2005 ... These innovations also served as the basis for the company's transfection formulations and nucleic acid labeling and ... Mirus Bio LLC (formerly Mirus Bio Corporation), develops and manufactures transfection reagents, electroporation solutions and ... new chemistries for transfection [9]; development of a non-viral vector providing sustained liver-specific transgene expression ...
Malone RW, Felgner PL, Verma IM (August 1989). "Cationic liposome-mediated RNA transfection". Proceedings of the National ...
Current transfection efficiency remains poor. One example of the successful use of L. monocytogenes in in vitro transfer ...
"Gene transfer (transfection) methods in animals , Genetic Engineering and Biotechnology Gene Transfer Methods and Transgenic ... or plants the process is called transformation and when it is used to deliver genes to animals it is called transfection. This ...
"Gene transfer (transfection) methods in animals , Genetic Engineering and Biotechnology Gene Transfer Methods and Transgenic ... Kim, Tae Kyung; Eberwine, James H. (August 2010). "Mammalian cell transfection: the present and the future". Analytical and ... so the process used to insert foreign DNA into animal cells is usually called transfection. There are many ways to directly ... this process is known as stable transfection. To create transgenic animals the DNA must be inserted into viable embryos or eggs ...
Single cell transfections are used to virtually transfer any type of mammalian cell into another using a syringe which creates ... Cuerrier, Charles M.; Lebel, Réjean; Grandbois, Michel (2007-04-13). "Single cell transfection using plasmid decorated AFM ...
"Stereoselective pH Responsive Peptide Dendrimers for siRNA Transfection". Bioconjugate Chemistry. 30 (8): 2165-2182. doi: ...
Moleleki N, van Heerden SW, Wingfield MJ, Wingfield BD, Preisig O (July 2003). "Transfection of Diaporthe perjuncta with ... to several fungal species that are closely related to Cryphonectria parasitica using in vitro virus transfection techniques. ...
The term can also be understood as DNA transfection using a viral vector. Viral transformation can occur both naturally and ... Transduction Transfection Transgenesis Baron, ed. by Samuel (1996). "Effects on Cells". Medical microbiology (4. ed.). ...
"Fluorescent Peptide Dendrimers for siRNA Transfection: Tracking pH Responsive Aggregation, siRNA Binding, and Cell Penetration ... "Stereoselective pH Responsive Peptide Dendrimers for siRNA Transfection". Bioconjugate Chemistry. 30 (8): 2165-2182. doi: ...
This could result in a PAMAM-DNA complex, which would make DNA transfection more efficient due to neutralization of the charges ... As with drug delivery applications, specific targeting of the transfection complex is ideal and must be explored as well. ... Although viral vectors can offer highly specific, high-efficiency transfection, the generation of such viruses is costly and ... Indeed, several reports have confirmed PAMAM dendrimers as effective DNA transfection agents. When the charge balance between ...
H. diminuta has an effective mechanism for interspecies transfection. Beetles prefer to ingest rat droppings infected with ...
Poly(ethylenimine) was the second polymeric transfection agent discovered, after poly-l-lysine. PEI condenses DNA into ... Akinc, A; Thomas, M; Klibanov, AM; Langer, R (2004). "Exploring polyethylenimine-mediated DNA transfection and the proton ... 2003). "Synthesis of Linear Polyethylenimine Derivatives for DNA Transfection". Bioconjugate Chemistry. 14 (3): 581-587. doi: ... for Transfection Purpose and Linear Pei Obtained with Such Method". Archived from the original on 2012-08-05. Steuerle, Ulrich ...
The use of particle bombardment, or the gene gun, is another physical method of DNA transfection. In this technique, DNA is ... This is the simplest method of non-viral transfection. Clinical trials carried out of intramuscular injection of a naked DNA ... Previously, low levels of transfection and expression of the gene held non-viral methods at a disadvantage; however, recent ... Inorganics have also been shown to exhibit improved in vitro transfection for attached cell lines due to their increased ...
Material intended for transfection is prepared similarly from high purity oleic acid. It was originally introduced into ... The pure material can also be used for the liposomal-transfection of DNA, RNA and other negatively charged molecules. The ... and Transfection;". Critical Reviews in Therapeutic Drug Carrier Systems. 21 (4): 257-317. doi:10.1615/ ...
She earned her PhD in 2010, "Optical sorting and photo-transfection of mammalian cells". She was a member of the SPIE students ... Optical sorting and photo-transfection of mammalian cells (Thesis). Dholakia, Kishan, Council for Scientific and Industrial ... "Graphene for improved femtosecond laser based pluripotent stem cell transfection". Journal of Biophotonics. 7 (5): 351-362. doi ...
... Transfection and Selection Data for HeLa Cells. *Rebecca Skloot, The Immortal Life of Henrietta Lacks book website with ...
Transfection is often accomplished by introducing vectors into a cell. Once internalized, the fusion protein nicks the target ...
Software features include time-lapse, cell counting and transfection analysis. A key feature of the EVOS fluorescence ...
Signal transduction Transfection - means of inserting DNA into a cell. Transformation (genetics) - means of inserting DNA into ... This plasmid is inserted (usually by transfection) into a producer cell together with other plasmids (DNA constructs) that ... so that simultaneous transfection of multiple plasmids is required to get infectious virions. Moreover, only the plasmid ...
With Graham's contributions in transfection of the cells with the adenovirus 5 this led to the cell line of HEK 293 cells. HEK ... The technique of calcium phosphate transfection became available in 1973. Fellow molecular biologist Piet Borst called this ... has performed research in adenoviruses and was fundamental in the creation of the technique of calcium phosphate transfection ...
Strategies for the Preparation of Synthetic Transfection Vec ... DNA Synthetic Transfection Vectors Targeted siRNA Transfection ... Strategies for the Preparation of Synthetic Transfection Vectors, by Asier Unciti-Broceta, Matthew N. Bacon, and Mark Bradley. ... Hyperbranched Polyamines for Transfection, by Wiebke Fischer, Marcelo Calderon, and Rainer Haag. * ... Chemically Programmed Polymers for Targeted DNA and siRNA Transfection, by Eveline Edith Salcher and Ernst Wagner. * ...
Transfection definition, the insertion into a cell of a bacterial plasmid that contains a foreign virus or genetic material. ... Words nearby transfection. transf., trans-fat, trans-fatty acid, transfd., transfect, transfection, trans female, transfeminine ...
GeneXPlus Transfection Reagent. GeneXPlus Transfection Reagent is designed to efficiently transfect a broad spectrum of cell ... TransfeX™ Transfection Reagent. TransfeX™ Transfection Reagent has been optimized for use on a wide range of cell types, ... Our transfection reagents are cationic lipid-based; due to their positive charge, these transfection reagents interact with the ... ATCC® Transfection reagents may be used for the transfer of nucleic acids for a variety of applications, including protein ...
... and merge transfection and expression steps in bioprocessing. ... New transfection technologies abound. They are making it ... Transformative Times for Transfection. No Longer Mere Methodology, Transfection Is Being Integrated into New Applications ... "The biggest challenge is to try to optimize transfection on microcarriers," notes Dr. Rancourt. Transfection in the stirred- ... "But when a transfection experiment is performed, the phenotype is a spectrum that depends on where the gene may integrate into ...
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TransFast™ Reagent is more versatile than many other traditional transfection methods. ... Transfection Reagents TransFast™ Transfection Reagent. Fast, Efficient Transfection. *Transfect in 1 hour (or as little as 30 ... ViaFect™ Transfection Reagent. High-efficiency, low-toxicity transfection of many cell types, including iPS cells. ... ProFection® Mammalian Transfection System. Calcium phosphate-based method for stable or transient transfection of eukaryotic ...
... common transfection methods, and the role of transfection in gene editing and silencing for research, drug discovery, ... Transfection. Transfection reagents, protocols, and resources including Roche X-tremeGENE transfection reagents for DNA, siRNA ... Introduction to Cell Transfection. Transfection is the introduction of DNA, RNA, or proteins into eukaryotic cells and is used ... Calcium Phosphate Transfection Kit Protocol Calcium phosphate transfection is a common method for the introduction of DNA into ...
... using various stock images to create a transfection vision)and Layout Design for the New Thermo Scientific Transfection Product ... Thermo Scientific Transfection Brochure & Cover Imagery. Created Cover Imagery (using various stock images to create a ... transfection vision)and Layout Design for the New Thermo Scientific Transfection Pr Read more. 7 ...
... kathy gately kathy.gately at Mon Jul 6 14:53:41 EST 1998 *Previous message: Smeared ... Hi, Has anybody transfected HepG2 cells with SV40-CAT or CMV enhanced tk-CAT? I have done several transfections and Ive used ...
Transfection,Reagent,Handbook,biological,advanced biology technology,biology laboratory technology,biology device technology, ... For transfection of eukaryotic cells with RNA and siRNA Contents Kit Contents Storage and Stability Quality Control a href #Te, ... Optimizing siRNA Transfection. Guidelines for Transfection of siRNA Duplexes Using TransMessenger Transfection Reagent. ... TransMessenger Transfection Reagent. 2. Efficient DNA transfection of primary CNS neurons using TransMessenger Transfection ...
When I used stock pcDNA-lacZ for , transfection, everything worked fine and b-gal was obviously detected by , western blot ... transfection problem: DNA purity. Bruno Cenni bruno.removethis.cenni at Mon Nov 19 02:11:57 EST 2001 * ... Ive a problem during transfection. I tried to transfect pcDNA-lacZ to , mammalian cells using lipofectamine plus. ...
Transfection of RNA can be used either to induce protein expression, or to repress it using antisense or RNA interference (RNAi ... procedures.,/p, ,p,There are two types of transfection - transient and stable - suited to different experimental applications, ... p,Transfection - the delivery of DNA or RNA into eukaryotic cells - is a powerful tool used to study and control gene ... Guidelines for transfection of DNA. Guidelines for transfection of RNA. Guidelines for transfection of siRNA. Performing ...
... a catheter arrangement with various embodiments for applying heat to a patients cells in vivo in order to improve transfection ... SMC transfection was performed using DNA coprecipitated with CaPO4. SMCs were immediately heated for up to 1 hour at 42 to 45 ... One technique for transfection of cells has used laser poration. This approach has been performed in vitro using a laser beam ... Other approaches to transfection of cells have included chemical methods or electrical poration used in a cell culture, but ...
R2 Transfection Reagent is a two component non-liposomal reagent combination developed specifically for efficient transfection ... siRNA Transfection Guidelines: *The transfection of siRNA must be optimal in order to obtain maximum silencing of a target gene ... The transfection efficiency of plasmic DNA varies from cell line to cell line. For efficient DNA transfection we recommend the ... In order to easily estimate the efficiency of transfection of particular cell lines use Fluorescein-siRNA Transfection Control ...
Transfection is the process of introducing certain nucleic acids into a eukaryotic cell by means other than a virus. While the ... Two main types of transfection exist: transient transfection and stable transfection. In transient transfection, the DNA is ... In stable transfections, the new DNA becomes part of the cells original DNA by either adding on to it or replacing a piece of ... Transfection is the process of introducing certain nucleic acids into a eukaryotic cell by means other than through a virus. ...
Or does it only matter at transfection? How do I know that my cells are in the mid log phase? How do i... ... confluent and preferably not in colonies for optimal transfection. I usually split my cells 24 hours before transfection. ... Should I always split my cells at their mid log phase? Or does it only matter at transfection? How do I know that my cells are ... Its all rough estimates, 80-100% means usually a full dish of cells, depending on the transfection methods and the cell lines ...
... Leif Søndergaard lsunicph at Thu Sep 20 08:16:17 EST 2001 *Previous message: Want More Money? ... efficiency of transfection, cell death etc. Has anyone used this ,method and got it to work consistently well? Alternatively, ...
Efficient expression of tetracycline-responsive gene following transfection of dentate gyrus neurons in vitro. ...
Reverse transfection method. US6951757 *. Mar 4, 2003. Oct 4, 2005. Whitehead Institute For Biomedical Research. Transfection ... in order to promote co-transfection of the host cells with at least two different target sequences. Co-transfection refers to ... the yeast may be of any species which are cultivable and in the transfection array can be maintained upon transfection. ... For instance, the transfection array can provide a library of secreted peptides, and the ability of a given peptide to induce ...
It is a unique cationic lipid that has been specifically optimized for the transfection of neuronal cells. ... NeuroPORTER Transfection Kit is the latest innovation in DNA transfection. ... NeuroPORTER Transfection Kit is a unique cationic lipid that has been specifically optimized for the transfection of neuronal ... Life Science > Molecular Biology > Cloning & Expression > Transfection Reagents > NeuroPORTER™ Transfection Kit Data ...
... a novel polyamine formulation for transient and/or stable transfections in a wide range of cell types. This transfection ... Gain the proven advantages of the GeneJammer Transfection Reagent, ... The GeneJammer Transfection Reagent is a novel polyamine formulation for transient and/or stable transfections in a wide range ... The GeneJammer Transfection Reagent is a recommended alternative product to the discontinued SatisFection Transfection Reagent ...
Clickable Poly(ionic liquids): A Materials Platform for Transfection.. Freyer JL1, Brucks SD1, Gobieski GS1, Russell ST1, ... The resulting TAC polymers are biocompatible and efficient transfection agents. This robust, efficient, and orthogonal click ...
Production of high-titer helper-free retroviruses by transient transfection. W S Pear, G P Nolan, M L Scott, and D Baltimore ... Production of high-titer helper-free retroviruses by transient transfection Message Subject (Your Name) has sent you a message ... The generation of high-titer, helper-free retroviruses by transient transfection has been achieved by using the highly ... infectious units/ml of supernatant within 72 hr after CaPO4-mediated transfection. A stringent assay for replication-competent ...
Topical tissue nano-transfection mediates non-viral stroma reprogramming and rescue. *Daniel Gallego-Perez1,2,3,4. na1, ... Figure 1: TNT mediates enhanced reprogramming factor delivery and propagation beyond the transfection boundary.. ... Gallego-Perez, D., Pal, D., Ghatak, S. et al. Topical tissue nano-transfection mediates non-viral stroma reprogramming and ... Flow micropillar array electroporation to enhance size specific transfection to a large population of cells *Yingbo Zu ...
However, gene transfection in macrophages is difficult. We have shown here that macrophages are more resistant to gene ... transfection compared with other cell types. To further develop an efficient gene delivery system for macrophages, we evaluated ... Macrophage transfection studies are crucial for understanding gene regulation and expression. ... Macrophage transfection studies are crucial for understanding gene regulation and expression. However, gene transfection in ...
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Transfection. For consistent success in your Transfection and silencing RNA applications (to inhibit and suppress target ... LIFE SCIENCES > Molecular Biology - Genetics > Transfection>LIFE SCIENCES > Molecular Biology - Genetics > Transfection. ... our chemical-transfection agent UptiFectin™OFF. This is a patented synthetic derivative of a natural compound designed ...
Through careful optimization--e.g. choosing the right transfection agent and transfection method--high levels of transfection ... Low transfection efficiency and low cell viability are the most frequent causes of unsuccessful gene silencing experiments. ... efficiency can be achieved ,Optimizing,siRNA,Transfection,for,RNAi,biological,advanced biology technology,biology laboratory ... siPORT Amine Transfection Agent. 1 ml. 4510. siPORT NeoFX Transfection Agent. 0.4 ml. 4511. siPORT NeoFX Transfection Agent. 1 ...
This technique is rapid, reliable, uses minimal amounts of reagent per transfection, and yields high transfection rates in a ... Transfection of cells in culture with cDNA constructs is a powerful tool in cell biology, but postmitotic cells, including ... This protocol results in efficient (20 to 70%, depending on cell type) transfection of myotubes, high levels of cDNA expression ... Muscle cell biologists may now perform experiments in mature myotubes rather than relying on transfection of myoblast cultures ...
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  • Transfection reagents introduce nucleic acid constructs into eukaryotic cells for protein synthesis and the overexpression of mutant DNA constructs. (
  • ATCC ® Transfection reagents may be used for the transfer of nucleic acids for a variety of applications, including protein expression and the overexpression of mutant genes. (
  • due to their positive charge, these transfection reagents interact with the negatively charged backbone of nucleic acids and cell membranes. (
  • These exceptional transfection reagents, combined with ATCC's technical know-how, will enable your transcriptional and translational gene studies. (
  • ATCC offers a line of transfection reagents tailored for effective nucleic acid transfer into a wide variety of cells for a multitude of applications. (
  • The QIAGEN Transfection Tools web site contains siRNA transfection protocols and the most up-to-date information and literature on QIAGEN reagents for RNAi. (
  • Some transfection protocols require serum-free conditions for optimal performance, since serum can interfere with many commercially available transfection reagents. (
  • In addition, we have compared NeuroPORTER reagent with other commercially available transfection reagents using Human NT2 Neuron Precursor Cells, Differentiated hNT Neurons, and C6 Glioma Cell Lines. (
  • siRNAs can be transiently transfected using commonly available transfection reagents. (
  • The siPORT Transfection II Kit contains samples of both of these reagents as well as positive and negative control siRNAs for protocol optimization. (
  • Located in the Parc d'Innovation at Illkirch close to the city and University of Strasbourg in Eastern France, the company has been producing and selling its proprietary range of transfection reagents and technologies since 2001. (
  • To complement and enhance the existing portfolio of its transfection solutions, Qiagen recently launched the TransFect Protocol Database and two new DNA transfection reagents: Attractene Transfection Reagent and NanoFect Transfection Reagent. (
  • The latest reagents use advanced technology to meet all DNA transfection needs. (
  • Home brew reagents are still the preferred choice of reagents for transfection by researchers all over the world. (
  • Dharmacon transfection reagents are designed specifically for small RNA transfection (DharmaFECT), co-transfection of plasmid and small RNA ( DharmaFECT Duo ), plasmid transfection ( DharmaFECT kb ) and self delivering siRNA for difficult to transfect cell types ( Accell siRNA ). (
  • One of four siRNA/microRNA specific formulations, DharmaFECT 2 is a chemically distinct alternative to one-size-fits-all transfection reagents to achieve high-efficiency silencing in more cell types. (
  • Altogen Biosystems specializing in the development of pre-optimized in vitro transfection reagents and in vivo delivery systems, transfection products and services . (
  • Nucleic acid transfection is a procedure used in nearly any cellular laboratory and the abundance of commercially available reagents has made this a seemingly simple endeavor. (
  • Cationic lipid-or polymer-based reagents are the most common means of transfection and suitable to most cell types. (
  • Transfection reagents couple a nucleic acid or an expression plasmid to a cationic lipid or polymer producing a liposome that interacts with the cell membrane and results in endocytosis of the molecule. (
  • In the case of siRNA delivery, a new novel technology exists, in which the siRNA is chemically modified to facilitate uptake into the cell without the need for transfection reagents, instruments, or viral vectors. (
  • Modified siRNAs achieve gene silencing in cells that had been beyond the reach of conventional RNAi methods due to toxicity from transfection reagents or undesirable viral responses. (
  • She has led product development for Novagen and EMD Biosciences (now EMD Millipore) in the areas of transfection reagents and expression vectors. (
  • Products include DNA vectors for cloning and expression, cell transfection reagents and cell culture tools, immobilized and soluble enzymes, products for genomics and proteomics research, numerous antibodies and recombinant proteins, superior fluorescence reagents and kits, affinity chromatography products, as well as general laboratory equipment. (
  • Although transfection reagents are commonly used to deliver gene-silencing oligonucleotides to cells in culture, a relatively new technique that does not require delivery vehicles has been gaining traction. (
  • Unfortunately, since most transfection reagents damage cells during transfection, simply using more of it is not an option. (
  • Xfect also outperforms popular competitor transfection reagents in Jurkat cells , which are notoriously difficult to transfect. (
  • We also offer FuGENE ® HD and DharmaFECT ® Duo Transfection Reagents, as well as RapidTrans™ chemically competent E. coli , which are supplied in a convenient 96-tube tray. (
  • Compatible with commonly used transfection reagents (e.g. (
  • G-FECT™ DNA Transfection reagents are a series of powerful DNA transfection reagents for the effective and reproducible transfection of DNA into mammalian cells. (
  • Three transfection reagents, Lipofectamine ® 2000, TransIT-PRO ® and linear 25 kDa polyethylenimine were evaluated for transient expression of enhanced green fluorescent protein in Chinese hamster ovary cells. (
  • At Booth 1753, Lonza's experts will demonstrate how the company's instruments, reagents and software can be integrated into robotic platforms for automated workflow solutions, enabling more effective transfection and endotoxin testing. (
  • I'm not sure what your endpoint is for the successful transfection e.g. which protein you are trying to overexpress, but you could perhaps try a few different transfection reagents and insert GFP or something else for the readout. (
  • If this is the case, you may simply want to thaw a new tube and start your cells again because you will likely come across the same problem with other transfection reagents. (
  • PromoCell's PromoFectin Transfection Reagents are ideal for highly effective and reliable transfection of a variety of cell types, including many hard-to-transfect cell lines, neuronal cells, sensitive primary cells and insect cells. (
  • All commercial transfection reagents can be associated with MA Lipofection Enhancer reagent by simple mixing in order to generate magnetic delivery systems. (
  • The studies reported here expand earlier studies which compared the differential expression of transcripts following transfection by either the FuGENE® 6 [1] or FuGENE® HD [2, 3] Transfection Reagents to another reagent on the market using either a different gene or a vector that did not contain the gene of interest. (
  • We increased the number of transfection reagents tested, looked at the effect of plating cell density, and measured effects on the transcriptome at earlier time points. (
  • FuGENE® HD Transfection Reagent was tested in all experiments, while reagents from other vendors (Reagent L, Reagent L2K, Reagent LX, Reagent LXP, Reagent LT-1, and Reagent E) were not used in each experiment. (
  • Transfection technologies and reagents are combined with nanoparticles for better efficacy in the above mentioned studies. (
  • as efficient transfection reagents both in vitro and in vivo [1-2]. (
  • GeneX Plus displays high transfection efficiency with low cytotoxicity in suspension cells such as HEK-293T/SF, THP-1, as well as difficult-to-transfect cell lines such as RAW 264.7 and SH-SY5Y. (
  • Please keep in mind that these are only guidelines, and the efficiency of transfection can be dependent on many different parameters, such as siRNA quality, cell type, passage number, and confluency of the cells at the time of transfection. (
  • The apparatus may be a catheter arrangement with various embodiments for applying heat to a patient's cells in vivo in order to improve transfection efficiency or application efficiency. (
  • A high efficiency transfection protocol employing a common polycationic lipid is described. (
  • The transfection efficiency was determined by direct staining for X-gal. (
  • Thus, a dramatic increase in transfection efficiency can be obtained by simply repeating transfection with the use of a common polycationic lipid. (
  • Felgner PL (1993) LipofectAMINE ™ reagent: a new, high efficiency polycationic liposome transfection reagent. (
  • efficiency of transfection, cell death etc. (
  • NeuroPORTER provides a fast, simple, and reliable system for DNA transfections and assures high efficiency, high viability results every time! (
  • High-efficiency gene transfection of macrophages by lipoplexes. (
  • Macrophage transfection was maximal at the DNA:LipofectAMINE:protamine ratio of 1:12:1 mu g/ml The LipofectAMINE formulation showed a 10-12-fold increase in transfection efficiency over DOTAP and a 4-5-fold increase over Lipofectin. (
  • Low transfection efficiency and low cell viability are the most frequent causes of unsuccessful gene silencing experiments. (
  • The LipoTAXI mammalian transfection reagent contains a unique, low-toxicity lipid formulation that has been tested with over 30 cell lines and has been shown to achieve high efficiency transfections with a wide variety of cells. (
  • Compared to CaPO4/DEAE-mediated transfection, it provides a 10-fold increase in transient transfection efficiency in many cell lines. (
  • The as-synthesized F-CDs achieves dramatic gene transfection efficiency as well as low cytotoxicity in commonly used cell lines at low weight ratio. (
  • The long-term goal of this project is to develop commercial products for delivering various biomolecules into mammalian cells with perfect transfection efficiency. (
  • It delivers exceptional transfection efficiency into the widest range of difficult-to-transfect and common cell types with improved cell viability. (
  • Calcium Phosphate Transfection Method This calcium phosphate transfection method works best in cell lines that are 1) highly transformed and 2) adherent (Hela, U2OS, SAOS2, AdAH, NPC-KT and obtain from 20% to 100% transfection efficiency depending on the cell line). (
  • Determining the optimal plating conditions for high transfection efficiency and low cytotoxicity can be highly involved. (
  • Using this novel system, a plasmid transfection efficiency up to ∼72% was obtained for CHO-K1 cells. (
  • In animal models there are additional factors to consider, including the presence of nucleases and the animal's innate inflammatory response, which can interfere with transfection efficiency. (
  • Toxicity and efficiency are the factors that have the biggest impact on the success of transfection experiments and can significantly affect important downstream applications. (
  • So far, viral vectors have been mainly used because of their inherently high transfection efficiency of gene. (
  • An obvious way to increase transfection efficiency is to increase the amount of plasmid DNA and transfection reagent mixture that you add to your cells. (
  • Xfect is a biodegradable transfection polymer, screened from a pool of 2,300 candidates for its very low cytotoxicity profile and high transfection efficiency. (
  • Xfect allows you to increase your transfection efficiency to the level you need. (
  • 40-fold higher transfection efficiency than Product L. This transfection reagent can also be used to transfect human adult stem cells. (
  • These defensive cellular strategies can greatly affect the efficiency of both transient and stable transfections. (
  • After 48 hours, transfection efficiency (% GFP-expressing cells) was measured using flow cytometry. (
  • It has several unique features necessary for efficient transfection - DNA condensation and endosomal release, which improves gene transfection efficiency. (
  • DNAfectamine Transfection Reagent comprises of four unique formulations of polycations and liposomes, which will guarantee high transfection efficiency and low cytotoxicity for any cell type including primary cells. (
  • For increased transfection efficiency. (
  • A modified version of G-FECT™-Plus that offers low cell toxicity and increased transfection efficiency. (
  • The goal of this study was to illustrate the versatility of Invitrogen Lipofectamine Stem Transfection Reagent, which was developed specifically for stem cells, to co-deliver multiple payloads in addition to large plasmid constructs with high transfection efficiency. (
  • Designed for dramatically improved transfection efficiency, Invitrogen Lipofectamine Stem Transfection Reagent offers a simple, robust and reproducible method for delivering DNA, RNA and RNP such as Cas9/gRNA complexes into a wide range of stem cells, including pluripotent stem cells, neural stem cells and mesenchymal stem cells (Figure 2) . (
  • This newest addition to the Lipofectamine transfection reagent family helps ensure high-efficiency transfection while maintaining maximum cell viability and growth in an undifferentiated state. (
  • Advantages of cationic lipid-mediated transfection include high efficiency, ability to transfect a wide range of cell types, reproducibility, low toxicity, and simplicity. (
  • If you let them reach confluency, even if this happened a few passages ago, the transfection efficiency will be significantly diminished. (
  • My point was that if you use cells that have reached 100% confluency your transfection efficiency will not be as good as when you use cells that have not reached 100% confluency. (
  • Of the lipoplexes prepared, those formulated with DSTAP showed higher transfection efficacy, however, the effect of buffer on transfection efficiency was formulation dependent. (
  • When MA Lipofection Enhancer is used, does the mixing order of components influence the final transfection efficiency? (
  • Depending on the transfection reagent used, the mixing order of components may influence the final transfection efficiency of MATra. (
  • The findings have been published in ( ACS Applied Materials & Interfaces , 'Effect of PEGylated Magnetic PLGA-PEI Nanoparticles on Primary Hippocampal Neurons: Reduced Nanoneurotoxicity and Enhanced Transfection Efficiency with Magnetofection' ). (
  • Furthermore, they found increased gene transfection efficiency in primary hippocampal neurons using PEGylated MNP-PLGA-PEI nanoparticles under an external magnetic field. (
  • Quantification of transfection efficiency. (
  • Effectene Reagent is used in conjunction with the enhancer and the DNA condensation buffer (Buffer EC) to achieve high transfection efficiency. (
  • The advantages of reverse transfection (over conventional transfection) are: The addition and attachment of target cells to the DNA-loaded surface can lead to a higher probability of cell-DNA contact, potentially leading to higher transfection efficiency. (
  • Exact-replicate arrays may be produced, since the same sample source plate may be dried and printed on different slides for at least 15 months' storage without apparent loss of transfection efficiency. (
  • To achieve the best results in siRNA transfection we recommend optimizing the following parameters. (
  • The amount of siRNA used is a critical factor for efficient transfection and gene silencing. (
  • The recommended starting amount for transfection of siRNA in 24-well plates is 0.8 g. (
  • A pipetting scheme for optimizing the transfection of siRNA in adherent cells in a 24-well format is provided in Table 4 on page 16. (
  • The ratio of TransMessenger Transfection Reagent to siRNA should be optimized for every new cell type and siRNA combination used. (
  • As a starting point for optimization we recommend using an siRNA to TransMessenger Transfection Reagent ratio (g:l) of 1:5 when using 24-well plates. (
  • 2). This procedure is provided as a starting point for optimization of siRNA transfection in mammalian cells using TransMessenger Transfection Reagent. (
  • On the day of transfection, dilute 1.6 l Enhancer R in Buffer EC-R. Add 0.8 g siRNA (minimum siRNA concentration, 0.1 g/l) and mix by vortexing. (
  • Transfection reagent is added directly to the diluted DNA or siRNA to produce reagent-NA complexes. (
  • The TransPass™ R2 Transfection Reagent is a two component non-liposomal reagent combination developed specifically for efficient transfection of siRNA in a variety of mammalian cell lines that include "difficult to transfect" cells such as primary cells, endothelial cells, lymphocytes, muscle cells, etc. (
  • HeLa cells were transfected with different concentrations of p38 MAPK ShortCut siRNA Mix (+) or 24 nM eGFP ShortCut siRNA Mix (C) using TransPass R2 Transfection Reagent. (
  • Effective silencing of p38 MAPK is achieved at low siRNA nanomolar concentration 48 hours post-transfection. (
  • Optimizing siRNA Transfection for RNAi ( Low transfection. (
  • These transfection parameters include culture conditions, choice and amount of transfection agent, exposure time of transfection agent to cells, and siRNA quantity and quality. (
  • There is no single transfection parameter that by itself ensures efficient siRNA uptake by cells in culture. (
  • Ambion provides a series of tools to simplify RNAi experiments including two transfection agents designed specifically for siRNA delivery. (
  • Ambion's siRNA Delivery Resource has further details on transfection optimization, recommendations for transfection agent, and conditions to use with specific cell types. (
  • This article summarizes the use of reverse transfection to maximize performance of siRNA in cultured cells and offers suggestions on how to optimize siRNA transfection parameters. (
  • The TransFect Database provides DNA, RNA, and siRNA transfection protocols for a wide range of cell types and plate or dish formats. (
  • The most broadly applicable DharmaFECT formulation for optimal siRNA or microRNA transfection into a wide range of cell types for successful RNAi experiments. (
  • An aliquot of each of the four DharmaFECT formulations for siRNA/microRNA transfection optimization studies. (
  • This in vivo reagent is designed for targeted siRNA delivery into pancreas, however it is also compatible for transfection of any negatively charged molecules including RNA, DNA, proteins, and small molecules. (
  • Pancreas-targeted siRNA In Vivo Transfection Kit enables researchers to utilize in vivo transfection method to introduce DNA, RNA, and other molecules into pancreas cells. (
  • The Span-8 pipettors and the selective tip feature of the multichannel head were used to serially dilute different lipids and combine them with two concentrations of a FAM-labelled siRNA (siGLO Green) which acted as a transfection readout. (
  • Common transfection examples include introducing DNA plasmids that have gene inserts for expression, or small interfering RNA (siRNA) for targeted gene silencing. (
  • The siRNA Transfection Reagent is provided as a sterile solution, and is compatible with serum and antibiotics. (
  • Lipofectamine 2000 Transfection Reagent is best transfection reagent available from Life Technologies for the co-transfection of siRNA and plasmid DNA. (
  • siTran 1.0 - For transfection of the siRNA duplex siTran 1.0 is a brand-new transfection reagent specially for siRNA duplexes in transient transfection, high-throughput screening, etc. siTran 1.0 is also highly effective in transfecting plasmid DNA. (
  • Viromers, a technology breakthrough, take advantage of a viral fusion mechanism (hence their name) and translate this into polymer chemistry for plasmid and siRNA transfection. (
  • Transfection of THP-1 cells with plasmids or small interfering RNA (siRNA) is achieved by electroporation using the Lonza Nucleofector technology (Basel, Switzerland). (
  • We recommend to use 1 µl of MA Lipofection Enhancer per µg of DNA in initial experiments (for antisense oligos or siRNA ratios need to be optimized) and to prepare the DNA/transfection reagent complexes according to the reagent's manufacturer instructions first and then add the nanoparticles. (
  • There are several factors that can influence successful transfections, e.g . viability and density of the cells, choice of the transfection reagent, quality and type of the transfected molecules (plasmids, siRNA, oligonucleotides), as well as the culture medium and supplements used. (
  • GeneX Plus Transfection Reagent is a high-performance, animal-origin free, broad spectrum reagent that provides exceptional transfection of plasmid DNA into mammalian cells. (
  • PEI is clearly the reagent of choice for research into large-scale mammalian transient transfection, and this agreement allows Kempbio to use PEI for its transfection services offering. (
  • Linear polyethylenimine (l-PEI), one of the most popular non-viral transfection agents for mammalian cells in general, only achieves transfection rates in the single digit percentage range for these cells. (
  • The Xfect Protein Transfection Reagent uses a cell-penetrating peptide developed at Clontech to bind and transport active proteins directly into a wide variety of mammalian cell types, including hard-to-transfect human suspension cell lines and mouse embryonic stem cells. (
  • Transfection - the transfer of genetic material into mammalian cells - is a valuable tool that allows scientists to genetically manipulate neurons and neuronal tissue. (
  • Here we describe methods for rapid synthesis of gRNA and for delivery of Cas9 protein/gRNA ribonucleoprotein complexes (Cas9 RNPs) into a variety of mammalian cells through liposome-mediated transfection or electroporation. (
  • TransfeX™ Transfection Reagent has been optimized for use on a wide range of cell types, including cells that are generally difficult to transfect, such as hTERT immortalized cell lines, primary cells, and stem cells. (
  • After stable transfection (electroporation) and G418 selection in 293-cells i got nice fluorescent clones suitable for FACS and confocal IF. (
  • Transfection is the process of introducing nucleic acids into eukaryotic cells. (
  • Transfection is the introduction of DNA, RNA, or proteins into eukaryotic cells and is used in research to study and modulate gene expression. (
  • Calcium phosphate transfection is a common method for the introduction of DNA into eukaryotic cells. (
  • In polymer-based transfection, exogenous DNA forms complexes with cationic polymers such as polyethylenimine (PEI) that enter host cells by endocytosis. (
  • This is achieved by counting cells prior to seeding and by keeping the time period between seeding and transfection (minimum 24 hours) constant. (
  • This will ensure that the cell density is not too high and that the cells are in optimal physiological condition on the day of transfection. (
  • Make sure that cells are in good condition and are seeded 24 h before transfection. (
  • Cells should be 5080% confluent on the day of transfection. (
  • Transfection - the delivery of DNA or RNA into eukaryotic cells - is a powerful tool used to study and control gene expression. (
  • Stably transfected cells can be selected by co-transfection of a second plasmid carrying an antibiotic-resistance gene or by providing a resistance gene on the same vector as the gene of interest. (
  • In fast-forward transfection, plating and transfection of cells are performed on the same day. (
  • Whereas, in a traditional transfection, cells are plated 24 hours prior to transfection. (
  • Cells are seeded in culture medium containing serum and antibiotics the day before transfection and incubated under normal growth conditions. (
  • During complex formation, the medium on the cells is changed, before the complexes are added to the cells (see figure Fast forward and reverse transfection ). (
  • In standard or fast-forward transfection, cells are added to plate wells first, followed by transfection complexes. (
  • Cells are added after complex formation in the wells, hence the term "reverse transfection" (see figure Steps in fast-forward and reverse transfection ). (
  • Using LipofectAMINE, a widely used transfection reagent, we transfected 293T cells with a plasmid harboring the β-galactosidase (β-gal) gene. (
  • Its all rough estimates, 80-100% means usually a full dish of cells, depending on the transfection methods and the cell lines used the general idea is that cells need to be around 50% confluent and preferably not in colonies for optimal transfection. (
  • I usually split my cells 24 hours before transfection. (
  • NeuroPORTER Transfection Kit is a unique cationic lipid that has been specifically optimized for the transfection of neuronal cells. (
  • Images represent transfection of plasmids expressing GFP into Rat Primary Cortical Neurons, C6 Glioma Cells, Human NT2 Cells, and Differentiated hNT Neurons. (
  • ated transfection procedure involves pre-plating cells, meaning the cells are allowed to attach, recover, and grow for 24 hours prior to transfection. (
  • Reverse transfection involves simultaneously transfecting and plating cells, much like procedures used for transfecting suspension cells. (
  • For maximal cell viability during transfection, cells must be healthy at the beginning of the experiment--healthy cells are easier to transfect than poorly maintained cells. (
  • Transfection of cells in culture with cDNA constructs is a powerful tool in cell biology, but postmitotic cells, including myotubes, can be hard to transfect with classic methods. (
  • Transfection is the process of deliberately introducing DNA or RNA into cells . (
  • Transfection can result in unexpected morphologies and abnormalities in target cells. (
  • Transfection with RNA molecules produces changes which cannot be permanently transmitted down a line of cells. (
  • Transfection of animal cells usually involves opening temporary pores (holes) in the cell membrane , to allow the cells to take up the vector. (
  • Suitable for transfection of Bone Marrow MSCs (PCS-500-012), Dermal Microvascular Endothelial cells (PCS-110-010), THP-1 cells (TIB-202), Raw 264.7 cells (TIB-71), and SH-SY5Y cells (CRL-2266). (
  • GeneX Plus Transfection Reagent has been optimized for use with HEK293T/17 SF suspension cells (ATCC ACS-4500) and HEK Plus SFM (ATCC ACS-4002) to reproducibly express a wide range of proteins at high levels. (
  • This transfection reagent may be used with both adherent cells and cells growing in suspension. (
  • Transfection protocols for adherent & suspension cells, β-Galactosidase activity test, β-Galactosidase assay, histochemical staining assay. (
  • Strasbourg, May 18, 2010 - Polyplus-transfection SA, a privately-held company developing innovative solutions for molecular and cellular biology, announced today that Kempbio, Inc., a company dedicated to providing high quality bioservices to the biotechnology and biopharmaceutical industry, has signed a license agreement to use polyethylenimine (PEI) for the transfection of cells used for the production of recombinant proteins. (
  • For example, unlike traditional transfection, with PULSin you can study lethal proteins and control the level and time course of proteins in cells. (
  • Attractene Reagent is a nonliposomal lipid which enables efficient DNA transfection of adherent cells, including difficult-to-transfect cell types such as HaCaT, MonoMac6, and HCT116. (
  • Unlike transient transfection, in which introduced DNA persists in cells for several days, stable transfection introduces DNA into cells long-term. (
  • Successful stable transfection requires both effective DNA delivery and a way to select cells that have acquired the DNA. (
  • One of the most reliable ways to select cells that stably express transfected DNA is to include a selectable marker on the DNA construct used for transfection or on a separate vector that is co-transfected into the cell, and then apply the appropriate selective pressure to the cells after a short recovery period. (
  • Transfection in hard to transfect cells is the need of hour and it is a major challenge. (
  • Transfection (the process of deliberately introducing genes into cells) technologies have been used in life science research and has been exploited for therapeutic delivery like for eg, gene therapy and electro-chemotherapy. (
  • Results were again significantly better than for l-PEI, although further research into the response of individual T cells to the transfection agent will be necessary, before either method can be used to routinely transfect primary T lymphocytes. (
  • LAS VEGAS , March 6, 2012 /PRNewswire/ -- Altogen Biosystems announced the launch of its new pancreas-targeted in vivo RNAi transfection system in the company's popular in vivo delivery product line for use in laboratories and research facilities for development of new medicines and testing in cells and animals. (
  • Transfection - the process of transferring genetic material into cells - is a powerful tool for the rapid and efficient manipulation of gene expression in cells. (
  • Xfect Protein Transfection Reagent makes it possible to deliver active proteins directly into cells for studies that involve transcriptional regulation, the cell cycle, apoptosis, oncogenesis, epigenetics, cell regeneration, and transdifferentiation. (
  • Xfect Protein Transfection Reagent offers the best advantages of both, i.e., it retains low cytotoxicity and delivers more protein to a higher percentage of target cells (see Images & Data tab). (
  • Moreover, Xfect Protein can transfect cells that are growing at a higher density than competing products, which is important because assays are often performed within a few hours post-transfection and the higher cell densities ensure sufficient material for downstream analysis. (
  • The unique properties of the delicate cells make transfection challenging and necessitate specialized techniques. (
  • This video will review the principles behind transfecting neurons and introduce strategies commonly used on these cells, including nucleofection, biolistic transfection, and viral transduction. (
  • Because most cultured neurons are post-mitotic - or non-dividing - cells, specialized transfection protocols are required in order to successfully induce gene expression. (
  • Calcium phosphate transfection method is a very efficient means of introducing DNA into cells in many cell systems. (
  • Calcium-phosphate-mediated Transfection of Cells with High-molecular-weight Genomic DNA This protocol,is based on the venerable method of Graham and van der Eb (1973). (
  • Calcium-phosphate-mediated Transfection of Eukaryotic Cells with Plasmid DNAs Calcium phosphate forms an insoluble precipitate with DNA, which attaches to the cell surface and is taken into the cells by endocytosis. (
  • 1996) who rigorously optimized calcium-phosphate-based transfection methods for Chinese hamster ovary cells and the 293 line of human embryonic kidney cells. (
  • The precise pH of the transfection medium and the incubation time of cells with the coprecipitate are critical for reproducible and efficient transfection. (
  • Differences can come from the transfection lipid chosen, the concentrations of the lipids and nucleic acids, and the number of cells plated. (
  • Optical transfection is a powerful method for targeted delivery of therapeutic agents to biological cells. (
  • Optimizing transfection protocols ensures maximal nucleic acid uptake by a majority of cells while minimizing cytotoxicity. (
  • Transfection of cells in culture is a key technique in the molecular and cell biology toolbox. (
  • Our customers have indicated via survey that they have successfully used Xfect Transfection Reagent with a wide variety of cells and cell lines . (
  • The new unit expands the proven 4D-Nucleofector™ System to closed, larger-scale transfection of up to 1x10 9 cells. (
  • When it was introduced in 2001, Nucleofector™ Technology was the first efficient, non-viral transfection method for primary cells and hard-to-transfect cell lines. (
  • We show that combined knockdown of interferon-β (Ifnb1), Eif2ak2, and Stat2 rescues cells from the innate immune response triggered by frequent long-RNA transfection. (
  • Gentle on cells with no toxicity in all tested transfection protocols. (
  • 95%) proprietary blend of innate immune system inhibitors and has been functionally tested in transfection assays using THP-1 and RAW 264.7 cells. (
  • Bottom - Transient transfections of an ~3 kb GFP-expressing plasmid (left) and an 8 kb GFP-expressing plasmid (right) into THP-1 cells was performed using commonly used transfection methods such as GeneXPlus, Lipofectamine® LTX, jetPRIME®, and nucleofection. (
  • GeneJuice Transfection Reagent provides excellent performance in both stable and transient transfection of eukaryotic cells and is ideal for use with the Novagen pTriEx™ expression vectors. (
  • A cationic liposome-forming compound used for transfection of DNA, RNA, and other negatively charged molecules into eukaryotic cells. (
  • TurboFectin is a new generation of transfection reagent for nucleic acid delivery into eukaryotic cells. (
  • MegaTran 1.0 is a polymer based transfection reagent specially designed and manufactured for high volume DNA transfection and large scale protein production in HEK293T, CHO and 293S cells. (
  • Herein, we develop an efficient mRNA transfection in cultured mouse cardiac fibroblasts via a polyarginine-fused heart-targeting peptide and lipofectamine complex, termed C-Lipo and demonstrate the partial direct reprogramming of cardiac fibroblasts towards cardiomyocyte cells. (
  • A knot polymer, poly[bis(2-acryloyl)oxyethyl disulphide- co -2-(dimethylamino) ethyl methacrylate] (DSP), was synthesized, optimized and evaluated as a non-viral vector for gene transfection for skin cells, keratinocytes. (
  • Experience superior transfection in stem cells for yourself. (
  • Stem cell transfection is a key technique for many researchers, but the sensitivity of stem cells has made them difficult to transfect efficiently using reagent-based protocols. (
  • In addition, Lipofectamine Stem reagent can be used for the transfection of mRNA for gene expression or directed differentiation studies using stem cells. (
  • Sou, S. , Polizzi, K. and Kontoravdi, C. (2013) Evaluation of transfection methods for transient gene expression in Chinese hamster ovary cells. (
  • Xie, Q.L., Guo, X.Y., Chen, X.J. and Wang, Y.Y. (2013) PEI/DNA formation affects transient gene expression in suspension Chinese hamster ovary cells via a one-step transfection process. (
  • I have been trying to transfect HEK293 cells with Transfast and Lipofectamine and have not been able to confirm successful transfection. (
  • Hi Ivan i have a question regarding confluency of HEK293 cells for transfection.Yous uggested to thaw a new vial of cells if the cells had reached confluency( even if it was few passages before). (
  • Does it mean that once HEK293 cells reached 100% confluency, it CANNOT be used at all for transfection expts? (
  • The β-Galactosidase Staining Kit is an easy-to-use and efficient method to determine the percentage of cells expressing lac Z following transient or stable transfection of plasmids that contain lac Z. (
  • These include the use of difficult-to-label target cells, or, regarding reporter gene transfection-based assays, the use of difficult-to-transfect targets such as primary human professional antigen presenting cells (APCs). (
  • Magnet Assisted Transfection (MATra) is an easy-to-handle and extremely fast technology to optimize transfection of cells in culture. (
  • Exploiting a strong magnetic force, the full nucleic acid dose bound to magnetic nanoparticles is rapidly drawn towards the target cells resulting in a very efficient transfection. (
  • Transfection is a method for delivering nucleic acids, either DNA or RNA, into animal cells in vitro. (
  • Briefly, after overnight incubation, MCF7 (ATCC® HTB-22TM) cells plated in 6-well plates at either 200,000 or 400,000 cells per well, were transfected using the optimized amounts for each transfection reagent. (
  • At two time points post transfection (24 and 48 hours), supernatant from the cells was diluted (1:400) and measured for SEAP activity, according to the protocol for the chemiluminescent SEAP Reporter Gene Assay Kit (Cat No. 11 779 842 001). (
  • At three time points post transfection (8, 24 and 48 hours), cells were harvested and further processed at Cogenics, a Division of Clinical Data. (
  • What would be the best transfection reagent for HepG2 cells? (
  • Albany, NY -- ( SBWIRE ) -- 01/16/2017 -- Transfection is a process that involves production of genetically modified cells with the delivery of foreign nucleic acid (DNA or RNA) into the cell. (
  • Suspension (A) or adherent cells (B) were transfected with pAAV-hrGFP, pAAV-RC, and pAAV-Helper (1:1:1 ratio) using the TransIT-VirusGEN transfection reagent (2:1, vol:wt) at the indicated volumes per culture vessel. (
  • 1997. Cationic phosphonolipids as non viral vectors for DNA transfection in hematopoietic cell lines and CD34+ cells. (
  • In animal cells, transfection is the preferred term as transformation is also used to refer to progression to a cancerous state (carcinogenesis) in these cells. (
  • Transfection of animal cells typically involves opening transient pores or "holes" in the cell membrane to allow the uptake of material. (
  • The original meaning of transfection was "infection by transformation", i.e., introduction of genetic material, DNA or RNA, from a prokaryote-infecting virus or bacteriophage into cells, resulting in an infection. (
  • Because the term transformation had another sense in animal cell biology (a genetic change allowing long-term propagation in culture, or acquisition of properties typical of cancer cells), the term transfection acquired, for animal cells, its present meaning of a change in cell properties caused by introduction of DNA. (
  • Reverse transfection is a technique for the transfer of genetic material into cells. (
  • As DNA is printed on a glass slide for the transfection process (the deliberate introduction of nucleic acids into cells) to occur before the addition of adherent cells, the order of addition of DNA and adherent cells is reverse that of conventional transfection. (
  • Thus, transfection techniques and protocols serve as an analytical tool that facilitates the characterization of genetic functions, protein synthesis, cell growth and development. (
  • The TransFect Protocol Database provides cell-specific transfection protocols. (
  • All transfection protocols are designed to enable genetic material to bypass this barrier. (
  • Transfection protocols can be established in smaller scale using the 4D-Nucleofector™ X Unit and smoothly transferred to larger scale without the need for re-optimization. (
  • By combining a Nucleofector™ Device with Lonza's easy-to-use Nucleofector™ Kits and the company's range of cell-type specific protocols, high transfection efficiencies can be reproducibly achieved with excellent cell viability. (
  • Successful transfection of THP-1 monocytes with subsequent phorbol 12-myristate 13-acetate (PMA)-induced differentiation into macrophages is not a trivial matter, because according to previous transfection protocols, cell viability is lost almost completely within 24 h of PMA treatment following transfection. (
  • MATra can also be adapted to high-throughput transfection assays using robotic stations and adapted protocols. (
  • We have tried a number of transfection protocols. (
  • In reverse-transfection, nucleic acid is added to plate wells, followed by transfection reagent. (
  • Both siPORT NeoFX and siPORT Amine Transfection Agents can be used for reverse transfection or standard transfection of siRNAs into a wide variety of cell types. (
  • Here, we show evidence that an alternative transfection procedure, termed reverse transfection [1] or neofection [2], offers several key benefits over the traditional pre-plating method. (
  • Schematic comparing forward and reverse transfection. (
  • Workflow comparison of forward transfection and reverse transfection, illustrating the labor- and reagent-saving features. (
  • The same-day transfection and plating or reverse transfection will also help boost your percent transfected. (
  • This transfection reagent is effective in both a serum-containing medium and serum-free medium, offering high efficiencies with minimal cytotoxicity. (
  • The principal obstacle for 'foreign' nucleic acids (i.e. plasmids) during eukaryotic cell transfection is their own detection by cytosolic DNA sensors of the innate immune system ( see 'Details' tab ). (
  • Lipofectamine Stem reagent offers expanded capabilities for stem cell transfection, making it possible to deliver DNA plasmids in sizes up to 11 kb. (
  • Lipofectamine Stem Reagent outperforms FuGENE HD Reagent in pluripotent stem cell transfection, delivering both small and large DNA plasmids, mRNA, and Cas9 protein complexes. (
  • Plasmid Transfection Medium Reduced-serum medium suitable for transfections with DNA Plasmids. (
  • If you are referring to our CRISPR/Cas9 KO Plasmid and HDR Plasmid co-transfection protocol (below) then the final volume is 150 µl after combining 1-3 µg of both plasmids. (
  • However, with the invention of different types of microarray printing systems, hundreds of transfection mixes (containing different DNA of interest) may be printed on the same slide for cell uptake of plasmids. (
  • Transfection is the process of introducing certain nucleic acids into a eukaryotic cell by means other than through a virus. (
  • The principal obstacle for 'foreign' nucleic acids (such as plasmid DNA) during eukaryotic cell transfection is their own detection by cytosolic nucleic acid sensors such as Absent in melanoma 2 (AIM2) and cyclic GMP-AMP synthase (cGAS) [1]. (
  • GeneX Plus Transfection Reagent is designed to efficiently transfect a broad spectrum of cell types. (
  • This technique is rapid, reliable, uses minimal amounts of reagent per transfection, and yields high transfection rates in a previously hard-to-transfect cell type. (
  • Virus-mediated transfection, also known as transduction, offers a means to reach hard-to-transfect cell types for protein overexpression or knockdown, and it is the most commonly used method in clinical research (Glover et al. (
  • G-FECT™ can be used for plasmid transfection, genome editing, virus production and more. (
  • Recommended for use with Plasmid Transfection Reagent (sc-108061) and suspension of DNA for transfection. (
  • But, used the Plasmid Transfection Medium with liposome transfection was useful for the primary cell lines. (
  • This transfection method showed minimal toxicity at the concentrations tested and was at least 20-25-fold superior to the most frequently used DEAE-dextran method for macrophage transfection. (
  • from non-animal sources, 293-Free Transfection Reagent gives minimal cellular toxicity. (
  • C-Lipo enabled the mRNA-induced direct cardiac reprogramming due to its efficient transfection with low toxicity, which allowed for multiple transfections of Gata4, Mef2c, and Tbx5 (GMT) mRNAs for a period of 2 weeks. (
  • To further develop an efficient gene delivery system for macrophages, we evaluated various liposomal and non-liposomal agents including LipofectAMINE(R), Lipofectin(R), DOTAP, DEAE-dextran, and the DNA condensing agent protamine sulfate for their ability to promote gene transfection. (
  • Lipofectamine® 3000 reagent leverages our most advanced lipid nanoparticle technology to enable superior transfection performance and reproducible results. (
  • Lipofectamine® 2000 has been the most-cited transfection reagent for over a decade. (
  • 1) Transfection lipids (Lipofectamine RNAiMAX (L), DharmaFECT 1-4, and negative control (0)), Opti-MEM media (light orange), and two siGLO Green concentrations (purple) were added to a 96-well plate. (
  • For gene editing, Lipofectamine Stem reagent allows the co-transfection of Cas9 protein complexed with guide RNAs along with single-stranded DNA for homology directed repair. (
  • As well as suggesting an 'Ultra' version of lipofectamine such as Lipofectamine LTX or plus, you could check that your detection method for detecting an efficient transfection is working properly. (
  • There are two types of transfection - transient and stable - suited to different experimental applications, and with different vector requirements. (
  • Two main types of transfection exist: transient transfection and stable transfection. (
  • The major types of Transfection include physical based (such as electroporation, nucleofection, magnetofection and gene gun), biochemical based(lipofection, calcium phosphate and dextran mediated) and viral based(adeno viruses and adeno-associated viruses based) transfection. (
  • The global transfection technologies market is concentrated with international and regional companies that offer products for different types of transfection methods for pharmaceutical and biotechnology companies, contract research organizations (CRO), and academic and research institutes. (
  • There are two types of transfection technologies that include stable and transient. (
  • MarketsandMarkets will host webinar to help you uncover the high growth opportunities in Global Transfection Technologies Market. (
  • Which applications/end-user segments are showing wider adoption of Global Transfection Technologies Market? (
  • In stable transfections, the new DNA becomes part of the cell's original DNA by either adding on to it or replacing a piece of the old DNA. (
  • The GeneJammer Transfection Reagent is a novel polyamine formulation for transient and/or stable transfections in a wide range of cell types. (
  • The health and viability of the cell line, quality of the nucleic acid used, transfection reagent, duration of transfection, and the presence or absence of serum can all play a part. (
  • The goal of transfection optimization is to determine the conditions that will provide maximum gene knockdown while maintaining an acceptable level of viability for the particular cell type (see sidebar, Two-Step Optimization Protocol ). (
  • 80% cell viability) at the optimal transfection concentration by MTT assay. (
  • This allows for high transfection efficiencies up to 99% and makes the transfection success independent from any cell proliferation. (
  • You should be able to get 80% transfection efficiencies with home made solutions for calcium phosphate transfections. (
  • Nucleic acids commonly used in transfection include DNA , RNA , and proteins, among other materials. (
  • Transfection is an enabler technology used for many cell based research activities with applications spanning production of recombinant proteins and recombinant cell lines, gene therapy, delivery of therapeutics and also drug discovery. (
  • Because this method can be used to silence the expression of specific proteins or to drive the expression of foreign or modified proteins, transfection is an extremely useful tool in the study of the cellular and molecular processes that govern neuron function. (
  • When using a conventional protein transfection reagent, transfected proteins are accumulated in endosome or lysosome, and most of such proteins are dissociated in lysosome proteolysis system. (
  • Protein transfection reagent has been widely used as it can deliver proteins faster than inducing proteins by gene transfer. (
  • Transfection & expression of recombinant DNA are frequently used to study the effects and regulation of genes and their encoded proteins. (
  • This agreement includes Polyethylenimine (PEI) reagent from Polypus Transfection S.A that will be utilized by the National Cancer Institute for in-vitro transfection exploration to produce transfected proteins, viruses and antibodies.Europe contributed the second largest share of the total market in 2012.Utilization of nanomedicine in diagnostics, targeted drug delivery, clinical trials and drug development studies is on the peak in Europe. (
  • Figure 2 shows the superior transfection and gene expression results obtained with the NeuroPORTER reagent. (
  • This reagent affords high levels of gene expression in a variety of cell types and is suitable for both transient and stable transfection. (
  • Protein transfection is extremely rapid compared to traditional gene expression studies using transfected DNA (1-2 hours compared to 18-48 hours), because it bypasses cellular processes such as transcription and translation. (
  • The data from the four transfection experiments were combined for gene expression profiling data analysis, as previously described [4]. (
  • Transfection of Hippocampal Neurons with Plasmid DNA Using Calcium Phosphate Coprecipitation This protocol describes transfection of plasmid DNA into primary hippocampal neurons using DNA/calcium-phosphate (CaPO4) coprecipitation. (
  • The reagent-based method is further segmented as lipid mediated transfection (Lipofection), calcium phosphate, cationic polymers, DEAE-dextran, activated dendrimers and magnetic beads. (
  • Transfection can be carried out using calcium phosphate (i.e. tricalcium phosphate), by electroporation, by cell squeezing or by mixing a cationic lipid with the material to produce liposomes that fuse with the cell membrane and deposit their cargo inside. (
  • This transfection technology performs the same tasks as other biochemical procedures utilizing polymers, DEAE-dextran, calcium phosphate, and electroporation. (
  • For cell types not amenable to lipid-mediated transfection, viral vectors are often employed. (
  • TransIT-VirusGEN® Transfection Reagent by Mirus, USA, is designed to enhance delivery of packaging and transfer vectors to adherent and suspension HEK 293 cell types to increase recombinant adeno-associated virus (AAV) and lentivirus production. (
  • These vectors were designed for use in germline transformation and cell culture transfection assays. (
  • TransIT-VirusGEN® is a high-performance transfection reagent for the reliable and efficient delivery of packaging and transfer vectors to adherent or suspension HEK293 cell types. (
  • Non-viral methods include physical methods such as electroporation, microinjection, gene gun, impalefection, hydrostatic pressure, continuous infusion, and sonication and chemical, such as lipofection, which is a lipid-mediated DNA-transfection process utilizing liposome vectors. (
  • Besides gelatin, atelocollagen and fibronetin are also successful transfection vectors for introducing foreign DNA into the cell nucleus. (
  • Step by step, using this technology, researchers have been able to bring more functionality to cell types that were considered to be refractory to transfection, such as primary neurons," states Dr. Berger. (
  • Jaworski J, Figiel I, Proszynski T, Kaczmarek L (2000) Efficient expression of tetracycline-responsive gene following transfection of dentate gyrus neurons in vitro. (
  • The first set of slides demonstrates the efficient transfection of a plasmid expressing GFP (green fluorescent protein) into a primary culture of rat cortical neurons. (
  • We describe here an alternative strategy using particle-mediated gene transfer for the transfection of neurons and glia in intact brain slices. (
  • Transfecting Cultured Hippocampal Neurons with an Actin-GFP Plasmid This protocol describes two transfection methods for expressing GFP-tagged actin in primary neurons. (
  • transfection of nucleic acids into primary neurons and cultured neuronal cell lines. (
  • To achieve maximum effectiveness of exogenously introduced siRNAs, transfection optimization experiments are required. (
  • Automated transfection optimization workflow. (
  • The Xfect transfection protocol requires few steps and minimal optimization. (
  • However, depending on the cell line to be transfected and the commercial transfection reagent used, the optimal composition may require the optimization of the MA Lipofection Enhancer/DNA ratio. (
  • Optimization experiments were performed for each transfection reagent according to manufacturer's instructions, prior to selecting the amount of transfection reagent to be used for the microarray experiments. (
  • With a wide variety of cationic lipids, Avanti would love to assist you with your liposomal transfection research. (
  • Viral based transfection. (
  • It is also important to note that the packaging cell line used for viral amplification needs to be transfected with a non-viral transfection method. (
  • Chemical-based transfection can be divided into several kinds: cyclodextrin, polymers, liposomes, or nanoparticles (with or without chemical or viral functionalization. (
  • Transfection of RNA can be used either to induce protein expression, or to repress it using antisense or RNA interference (RNAi) procedures. (
  • We are truly delighted to sign this license agreement with Kempbio, a company offering quality cell culture and protein expression services," said Mark Bloomfield, CEO of Polyplus-transfection. (
  • Protein/antibody delivery using PULSin may be superior to transfection in some applications. (
  • What is the Xfect Protein Transfection Reagent and how does it work? (
  • Xfect Protein Transfection Reagent is a modified peptide with cell-penetrating activity whose amino acid composition enables it to interact with a protein cargo and transport this protein across a cell membrane barrier. (
  • What are the advantages of Xfect Transfection Reagent compared to other protein delivery technologies? (
  • Further, we found that the off-target cleavage rate is reduced using Cas9 protein when compared to plasmid DNA transfection. (
  • As a result, repeated transfection with protein-encoding RNA causes cell death. (
  • Conclusions/Significance Our results demonstrate that suppressing innate immunity enables frequent transfection with protein-encoding RNA. (
  • With recessive dystrophic epidermolysis bullosa keratinocytes (RDEBK-TA4), the DSP exhibited high transfection efficacy with both Gaussia luciferase marker DNA and the full length COL7A1 transcript encoding the therapeutic type VII collagen protein (C7). (
  • Cell transfection promotes the rapid advancement of our knowledge of genetic regulation and protein function. (
  • This is not (usually) a problem if the vector carries a gene for a nontoxic protein and the vector stock has been freshly prepared by transfection. (
  • Transfection technology application market is further segmented as biomedical research, therapeutic delivery and protein production. (
  • Rising prevalence of various cancers (prostate, breast and lung), cardiovascular diseases (arrhythmias, ischemic heart disease) and growing mass protein production further accentuates the global transfection technology market. (
  • Transfection technologies are widely used in cell research, recombinant protein production, drug discovery, gene therapy and therapeutics. (
  • Polyplus-transfection SA is a biotechnology company researching, developing, manufacturing and marketing innovative solutions for scientists working in molecular and cell biology. (
  • Polyplus-transfection recently extended its product offering to molecular biologists with the launch and commercialization of ZNA (TM) modified oligonucleotides. (
  • Rated 4 out of 5 by Wind from High effciency Conventional liposome transfection may perform well for the robust cell lines, but the primary cell lines was not. (
  • Lipofection (or liposome transfection) is a technique used to inject genetic material into a cell by means of liposomes, which are vesicles that can easily merge with the cell membrane since they are both made of a phospholipid bilayer. (
  • Successful transfection is influenced by many factors. (
  • Apart from the cell type, successful transfection also depends on the culture's age and density at transfection, the vector used, the purity of the nucleic acids, and the experimental conditions. (
  • While a virus has the potential to transmit these foreign items through a cell's membrane and into the cell, transfection uses methods other than a virus for transmission. (
  • Our finding also opens the door for the development of reprogramming and directed-differentiation methods based on long-RNA transfection. (
  • Reagent based transfection technology market is experiencing significant growth owing to cost effectiveness and less complicated methods involved during transfection. (
  • North America is found dominating the market, that is driven by factors such as increasing cases of infectious diseases demanding the need of antibiotics and the growing use of cell-based products which require transfection methods to treat chronic diseases such as cancer, tuberculosis, and cardiovascular diseases. (
  • However, cytotoxic effects, technological complications associated with instrument-based and virus-based methods might restrict the demand of the global transfection technology market to a certain level. (
  • Transient transfection is most efficient when supercoiled plasmid DNA is used. (
  • The resulting TAC polymers are biocompatible and efficient transfection agents. (
  • Deterministic transfection drives efficient nonviral reprogramming and uncovers reprogramming barriers. (
  • This protocol results in efficient (20 to 70%, depending on cell type) transfection of myotubes, high levels of cDNA expression in individual myotubes, and little cellular damage. (
  • Furthermore, the F-CDs shows excellent efficient transfection even at low pDNA dose, low F-CDs transfection reagent dose and high serum concentration, indicating potential clinical applications. (
  • NanoFect Transfection Reagent uses advanced nanopolymer technology for highly efficient DNA transfection. (
  • This constitutes a particular promising approach for efficient transient transfection at large scale. (
  • Testing novel compounds require efficient and targeted delivery of pharmaceuticals into pancreas tissue (known as in vivo transfection ). (
  • With this new addition, small- and large-scale transfection applications are now united in one system based on the highly efficient and established Nucleofector™ Technology. (
  • Improved protocol for efficient nonviral transfection of premature THP-1 macrophages. (
  • High levels of dystrophin cDNA expression, and an efficient distant transfection effect with preferential intranuclei inclusion of MF-2 vehicle, are very encouraging for the development of a new constructive strategy in gene therapy trials of DMD. (
  • We report here for the first time efficient transfection of mdx mice myofibers with dystrophin gene constructs delivered in vivo by the microsphere particles MF-2. (
  • there is a growing demand for new transfection technologies to address unmet needs for therapeutic delivery which is driving the transfection market. (
  • Growth in gene therapy is also driving the transfection market. (
  • Each of the formulations was prepared by hydration in dH 2 O or phosphate buffer saline (PBS) to investigate the effect of buffer salts on lipoplex physicochemical characteristics and in vitro transfection. (
  • Visit the Transfection Tools site for answers to your RNAi questions . (
  • Failure to optimize critical transfection parameters can render RNAi effects undetectable in cell culture. (
  • There are few upcoming transfection technologies like nucleofection and magnetofection which are an off-shoot of electroporation, which have the potential to address these challenges but are currently cost prohibitive due to which the technologies may have a much slower uptake in the developing world. (
  • Lonza's Nucleofector™ Technology is an improved electroporation technique that has transformed transfection and allowed scientists achieve results never before possible. (
  • Experiments involving large constructs have typically been carried out using electroporation, as reagent-based transfection yielded poor results. (
  • Since the reagent is composed of animal-origin free components and is serum compatible, there is no need for any culture medium change after transfection. (
  • For ease of use, plasmid transfections can be carried out entirely in the presence of serum . (
  • Expressfect™ Transfection Reagent is a new generation cationic polymer reagent that is easy to handle, exhibits higher stability, and shows higher resistance to serum in cell culture media. (
  • Promote transfection using lipofection and support growth of HEK 293 cultures with GE Healthcare HyClone SFM4Transfx-293 media, a serum-free, animal-derived component free (ADCF) media. (
  • MATra transfections can be performed in the presence or absence of serum and/or antibiotics. (
  • This nanopackaged t-PA gene plasmid was further cross-linked to ultrasonic microbubbles prepared with sucrose and bovine serum albumin to form the ultrasonic-targeted agent for t-PA gene transfection. (
  • Even though mRNA delivery to cardiac fibroblasts has the therapeutic potential, mRNA transfection in cardiac fibroblasts has been challenging. (
  • GMT mRNA transfection of cultured mouse cardiac fibroblasts via C-Lipo significantly increased expression of the cardiomyocyte marker genes, Actc1 , Actn2 , Gja1 , Hand2 , and Tnnt2 , after 2 weeks of transfection. (
  • The generation of high-titer, helper-free retroviruses by transient transfection has been achieved by using the highly transfectable 293T cell line into which are stably introduced constructs that express retroviral packaging functions. (
  • In this webinar, we discuss the key steps and protocol considerations for performing high-quality transfections on a consistent basis, taking into consideration variables such as cell type, culture conditions, reagent concentrations, and other critical parameters. (
  • Transfection signals from native CAT-encoding DNA compared well with those from a synthetic DNA sequence adapted to the P. falciparum major codon bias, demonstrating effective expression of the bacterial sequence despite its use of rare P. falciparum codons. (
  • Macrophage transfection studies are crucial for understanding gene regulation and expression. (
  • Muscle cell biologists may now perform experiments in mature myotubes rather than relying on transfection of myoblast cultures or heterologous expression systems. (
  • See RheoSwitch® Inducible Expression System Manual (NEB #E3000) for transient transfection protocol using pNEBR-X1GLuc (NEB #N8080). (
  • Furthermore, the sensing of dsDNA by cGAS induces the production of type I interferons (IFNs), through the STING/TBK1/IRF3 signaling axis, as well as the expression of a number of interferon-stimulated genes (ISGs), which exert potent effector functions, which are highly unwanted in transfection [1]. (
  • This is true for most of the reagent based transfection reactions. (
  • Market Research Report Search Engine Added 'Transfection Technology Market (Reagent-based method, Instrument-based method and Virus-based method) - Global Industry Analysis, Size, Share, Growth, Trends and Forecast, 2013 - 2019' to its database. (
  • The transfection of erythrocyte-stage P. falciparum parasites advances our ability to pursue genetic analysis of this major pathogen. (
  • Alternatively, the gene gun technique blasts through transfection barriers, by shooting bullets carrying genetic material through both cell and nuclear membranes. (
  • The key growth area of transfection products is the application in therapeutic area, where transfection is effectively used in treatment of cancer and other genetic disorders. (