Puromycin: A cinnamamido ADENOSINE found in STREPTOMYCES alboniger. It inhibits protein synthesis by binding to RNA. It is an antineoplastic and antitrypanosomal agent and is used in research as an inhibitor of protein synthesis.Puromycin Aminonucleoside: PUROMYCIN derivative that lacks the methoxyphenylalanyl group on the amine of the sugar ring. It is an antibiotic with antineoplastic properties and can cause nephrosis.Nephrosis: Pathological processes of the KIDNEY without inflammatory or neoplastic components. Nephrosis may be a primary disorder or secondary complication of other diseases. It is characterized by the NEPHROTIC SYNDROME indicating the presence of PROTEINURIA and HYPOALBUMINEMIA with accompanying EDEMA.Ribosomes: Multicomponent ribonucleoprotein structures found in the CYTOPLASM of all cells, and in MITOCHONDRIA, and PLASTIDS. They function in PROTEIN BIOSYNTHESIS via GENETIC TRANSLATION.Cycloheximide: Antibiotic substance isolated from streptomycin-producing strains of Streptomyces griseus. It acts by inhibiting elongation during protein synthesis.Dactinomycin: A compound composed of a two CYCLIC PEPTIDES attached to a phenoxazine that is derived from STREPTOMYCES parvullus. It binds to DNA and inhibits RNA synthesis (transcription), with chain elongation more sensitive than initiation, termination, or release. As a result of impaired mRNA production, protein synthesis also declines after dactinomycin therapy. (From AMA Drug Evaluations Annual, 1993, p2015)Sparsomycin: An antitumor antibiotic produced by Streptomyces sparsogenes. It inhibits protein synthesis in 70S and 80S ribosomal systems.Nephrotic Syndrome: A condition characterized by severe PROTEINURIA, greater than 3.5 g/day in an average adult. The substantial loss of protein in the urine results in complications such as HYPOPROTEINEMIA; generalized EDEMA; HYPERTENSION; and HYPERLIPIDEMIAS. Diseases associated with nephrotic syndrome generally cause chronic kidney dysfunction.Protein Synthesis Inhibitors: Compounds which inhibit the synthesis of proteins. They are usually ANTI-BACTERIAL AGENTS or toxins. Mechanism of the action of inhibition includes the interruption of peptide-chain elongation, the blocking the A site of ribosomes, the misreading of the genetic code or the prevention of the attachment of oligosaccharide side chains to glycoproteins.Kidney Glomerulus: A cluster of convoluted capillaries beginning at each nephric tubule in the kidney and held together by connective tissue.Podocytes: Highly differentiated epithelial cells of the visceral layer of BOWMAN CAPSULE of the KIDNEY. They are composed of a cell body with major CELL SURFACE EXTENSIONS and secondary fingerlike extensions called pedicels. They enwrap the KIDNEY GLOMERULUS capillaries with their cell surface extensions forming a filtration structure. The pedicels of neighboring podocytes interdigitate with each other leaving between them filtration slits that are bridged by an extracellular structure impermeable to large macromolecules called the slit diaphragm, and provide the last barrier to protein loss in the KIDNEY.Emetine: The principal alkaloid of ipecac, from the ground roots of Uragoga (or Cephaelis) ipecacuanha or U. acuminata, of the Rubiaceae. It is used as an amebicide in many different preparations and may cause serious cardiac, hepatic, or renal damage and violent diarrhea and vomiting. Emetine inhibits protein synthesis in EUKARYOTIC CELLS but not PROKARYOTIC CELLS.Protein Biosynthesis: The biosynthesis of PEPTIDES and PROTEINS on RIBOSOMES, directed by MESSENGER RNA, via TRANSFER RNA that is charged with standard proteinogenic AMINO ACIDS.Chloramphenicol: An antibiotic first isolated from cultures of Streptomyces venequelae in 1947 but now produced synthetically. It has a relatively simple structure and was the first broad-spectrum antibiotic to be discovered. It acts by interfering with bacterial protein synthesis and is mainly bacteriostatic. (From Martindale, The Extra Pharmacopoeia, 29th ed, p106)Leucine: An essential branched-chain amino acid important for hemoglobin formation.Peptide Biosynthesis: The production of PEPTIDES or PROTEINS by the constituents of a living organism. The biosynthesis of proteins on RIBOSOMES following an RNA template is termed translation (TRANSLATION, GENETIC). There are other, non-ribosomal peptide biosynthesis (PEPTIDE BIOSYNTHESIS, NUCLEIC ACID-INDEPENDENT) mechanisms carried out by PEPTIDE SYNTHASES and PEPTIDYLTRANSFERASES. Further modifications of peptide chains yield functional peptide and protein molecules.TritiumProteinuria: The presence of proteins in the urine, an indicator of KIDNEY DISEASES.Peptide Chain Elongation, Translational: A process of GENETIC TRANSLATION, when an amino acid is transferred from its cognate TRANSFER RNA to the lengthening chain of PEPTIDES.Polyribosomes: A multiribosomal structure representing a linear array of RIBOSOMES held together by messenger RNA; (RNA, MESSENGER); They represent the active complexes in cellular protein synthesis and are able to incorporate amino acids into polypeptides both in vivo and in vitro. (From Rieger et al., Glossary of Genetics: Classical and Molecular, 5th ed)Carbon Isotopes: Stable carbon atoms that have the same atomic number as the element carbon, but differ in atomic weight. C-13 is a stable carbon isotope.Peptidyl Transferases: Acyltransferases that use AMINO ACYL TRNA as the amino acid donor in formation of a peptide bond. There are ribosomal and non-ribosomal peptidyltransferases.RNA, Transfer, Amino Acyl: Intermediates in protein biosynthesis. The compounds are formed from amino acids, ATP and transfer RNA, a reaction catalyzed by aminoacyl tRNA synthetase. They are key compounds in the genetic translation process.Phenylalanine: An essential aromatic amino acid that is a precursor of MELANIN; DOPAMINE; noradrenalin (NOREPINEPHRINE), and THYROXINE.Viomycin: A strongly basic peptide, antibiotic complex from several strains of Streptomyces. It is allergenic and toxic to kidneys and the labyrinth. Viomycin is used in tuberculosis as several different salts and in combination with other agents.RNA, Transfer: The small RNA molecules, 73-80 nucleotides long, that function during translation (TRANSLATION, GENETIC) to align AMINO ACIDS at the RIBOSOMES in a sequence determined by the mRNA (RNA, MESSENGER). There are about 30 different transfer RNAs. Each recognizes a specific CODON set on the mRNA through its own ANTICODON and as aminoacyl tRNAs (RNA, TRANSFER, AMINO ACYL), each carries a specific amino acid to the ribosome to add to the elongating peptide chains.RNA, Long Noncoding: A class of untranslated RNA molecules that are typically greater than 200 nucleotides in length and do not code for proteins. Members of this class have been found to play roles in transcriptional regulation, post-transcriptional processing, CHROMATIN REMODELING, and in the epigenetic control of chromatin.IllinoisGift Giving: The bestowing of tangible or intangible benefits, voluntarily and usually without expectation of anything in return. However, gift giving may be motivated by feelings of ALTRUISM or gratitude, by a sense of obligation, or by the hope of receiving something in return.RNA, Untranslated: RNA which does not code for protein but has some enzymatic, structural or regulatory function. Although ribosomal RNA (RNA, RIBOSOMAL) and transfer RNA (RNA, TRANSFER) are also untranslated RNAs they are not included in this scope.ChicagoRecombinant Proteins: Proteins prepared by recombinant DNA technology.Th1 Cells: Subset of helper-inducer T-lymphocytes which synthesize and secrete interleukin-2, gamma-interferon, and interleukin-12. Due to their ability to kill antigen-presenting cells and their lymphokine-mediated effector activity, Th1 cells are associated with vigorous delayed-type hypersensitivity reactions.Th2 Cells: Subset of helper-inducer T-lymphocytes which synthesize and secrete the interleukins IL-4, IL-5, IL-6, and IL-10. These cytokines influence B-cell development and antibody production as well as augmenting humoral responses.MicroRNAs: 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.Cytokines: Non-antibody proteins secreted by inflammatory leukocytes and some non-leukocytic cells, that act as intercellular mediators. They differ from classical hormones in that they are produced by a number of tissue or cell types rather than by specialized glands. They generally act locally in a paracrine or autocrine rather than endocrine manner.Mice, Inbred C57BLMolecular Sequence Data: 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.Interferon-gamma: The major interferon produced by mitogenically or antigenically stimulated LYMPHOCYTES. It is structurally different from TYPE I INTERFERON and its major activity is immunoregulation. It has been implicated in the expression of CLASS II HISTOCOMPATIBILITY ANTIGENS in cells that do not normally produce them, leading to AUTOIMMUNE DISEASES.Carcinoma, Squamous Cell: A carcinoma derived from stratified SQUAMOUS EPITHELIAL CELLS. It may also occur in sites where glandular or columnar epithelium is normally present. (From Stedman, 25th ed)Lung Neoplasms: Tumors or cancer of the LUNG.Glypicans: A family of GLYCOSYLPHOSPHATIDYLINOSITOL-anchored, cell-surface heparan sulfate proteoglycans that may play a role in CELL GROWTH PROCESSES and CELL DIFFERENTIATION by modulating ligand-receptor interactions.Lung: Either of the pair of organs occupying the cavity of the thorax that effect the aeration of the blood.Carcinoma, Hepatocellular: 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.Syndecans: A family of transmembrane glycoproteins that contain a short cytoplasmic domain, a single-span transmembrane domain, and an extracellular domain with heparin sulfate and CHONDROITIN SULFATE chains. Syndecans interact with a variety of heparin-binding INTERCELLULAR SIGNALING PEPTIDES AND PROTEINS and may play a role in modulating cellular signaling during EMBRYONIC DEVELOPMENT, tumorigenesis, and angiogenesis.Cell Proliferation: All of the processes involved in increasing CELL NUMBER including CELL DIVISION.Lentivirus: A genus of the family RETROVIRIDAE consisting of non-oncogenic retroviruses that produce multi-organ diseases characterized by long incubation periods and persistent infection. Lentiviruses are unique in that they contain open reading frames (ORFs) between the pol and env genes and in the 3' env region. Five serogroups are recognized, reflecting the mammalian hosts with which they are associated. HIV-1 is the type species.Gene Knockdown Techniques: 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.Public-Private Sector Partnerships: An organizational enterprise between a public sector agency, federal, state or local, and a private sector entity. Skills and assets of each sector are shared to deliver a service or facility for the benefit or use of the general public.RNA Interference: 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.RNA, Small Interfering: 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.Gene Library: A large collection of DNA fragments cloned (CLONING, MOLECULAR) from a given organism, tissue, organ, or cell type. It may contain complete genomic sequences (GENOMIC LIBRARY) or complementary DNA sequences, the latter being formed from messenger RNA and lacking intron sequences.Private Sector: That distinct portion of the institutional, industrial, or economic structure of a country that is controlled or owned by non-governmental, private interests.
It contains resistance genes against ampicillin and puromycin. Release 1 of the TRC lentiviral shRNA libraries consist of about ... Verified RNAi clones and entire libraries are made available both by Sigma-Aldrich and Open Biosystems. A set of candidate ... for 15'000 human and 15'000 mouse genes. These libraries should help the scientific community to analyse gene function by RNAi ... hairpins are selected based on the 1st Refseq transcript from each NCBI gene. They should be 21mers, be at least 25bp from ...
Resistance to puromycin is conferred by the pac gene encoding a puromycin N-acetyl-transferase (PAC) that was found in a ... N verify (what is Y. N ?) Infobox references Puromycin is an antibiotic protein synthesis inhibitor which causes premature ... Puromycin resistance in yeast can also be conferred through expression of the puromycin N-acetyl-transferase (pac) gene.[8] ... Puromycin-resistant transformants are selected in LB agar medium supplemented with 125 µg/ml of puromycin. But use of puromycin ...
In another study, hypoxia led to induction of genes associated with apoptosis and cell death, but repression of genes was not ... While studying the effect of proteoglycan composition on nephritic permselectivity, it was noted that puromycin treatment of ... heparan sulfate proteoglycan expression has been verified, although no distinction between syndecan and perlecan has been made ... In yet another mouse knock-in model, the perlecan gene was mutated by homologous recombination of the endogenous perlecan gene ...
The two genes are believed to result from a gene duplication event; the ancestral gene is estimated to be about 500 million ... verify). Oxytocin (Oxt; /ˌɒksɪˈtoʊsɪn/) is a peptide hormone and neuropeptide. Oxytocin is normally produced by the ... and puromycin have been found to inhibit the enzymatic degradation of oxytocin, though they also inhibit the degradation of ... Sausville E, Carney D, Battey J (August 1985). "The human vasopressin gene is linked to the oxytocin gene and is selectively ...
The difference in size may be due to the elimination of obsolete genes or the takeover of its function by nuclear genes. ... verify). Spectinomycin, sold under the tradename Trobicin among others, is an antibiotic useful for the treatment of gonorrhea ... It is present in the genome or plastome of many plastids as the spc operon, which is usually two to 10 genes long. ...
verify). Amikacin is an antibiotic used for a number of bacterial infections.[2] This includes joint infections, intra- ... is due to a mutation in the rrs gene, which codes for the 16S rRNA. Mutations such as these reduce the binding affinity of ...
Neamine is made from six genes, DOIS gene (btrC, neo7); L-glutamine:DOI aminotransferase gene (btrS, neo6); a putative ... verify). Neomycin is an aminoglycoside antibiotic found in many topical medications such as creams, ointments, and eyedrops. ... Neomycin resistance is conferred by either one of two aminoglycoside phosphotransferase genes.[9] A neo gene is commonly ... gene (btrB, neo18); a putative alcohol dehydrogenase gene (btrE, neo5); another putative dehydrogenase (similar to chorine ...
The transgene was visualized using a probe recognizing a 1-kb sequence in the puromycin resistance gene cassette. The pigs were ... We used DNA transposition in HepG2 human hepatocellular carcinoma cells to verify the functional integrity of the transposon- ... puromycin resistance gene; pA, polyadenylation signal; HAAT, human α1-antitrypsin promoter. (B) The function of the D374Y-PCSK9 ... The transposon also contained a puromycin resistance gene for selection. ...
Gene Targeting.. Hct116 Fbw7−/− gene targeting has been previously described, and DLD1 Fbw7-null cells were made using the same ... Cells were transduced with lentiviruses and were selected with puromycin, and clones were isolated by limiting dilution. Fbw7 ... methods (14). All clones were verified by Southern blotting, PCR, and genomic sequencing. Hct116 Fbw7+/R505C cells and LoVo ... Fbw7 mutations increase gene expression of mitochondrial signature genes. (A) Schema depicting the generation of isogenic ...
After 48 h, 2 μg/mL puromycin (Sigma Inc, USA) was used to select stably cells expressing desired gene or shRNA. ... Authentication of cells was verified by short tandem repeat DNA profiling.. Lentiviral particles were prepared as described ... Integrated analyses of gene expression profiles and ChIP-seq data. The effects of p53-R273 on gene expression was analyzed by ... Notably, among these six genes, KLF6 is a documented tumor suppressor protein that transactivates E-cadherin gene expression13 ...
Analysis of gene expression correlation between SERPINB9 and IFN signatures in TCGA data. TCGA gene expression data (RNA-Seq V2 ... Transfected tumor cells were selected with puromycin (1.5 μg/mL) for 10-14 days. The efficiency of knockdown was assessed using ... 62). The plasmid cloning was verified by sequencing using the U6 primer (5′-GAGGGCCTATTTCCCATGATTCC-3′). Cells were then ... Following the amplification, the Ct values for target genes and the reference gene, β-actin, were recorded. Fold induction was ...
The DNA sequence of this gene was modified by optimizing the codon usage and reducing CpG motifs to 16 without changing the ... Resistance to the antibiotic puromycin is conferred by the Pac gene encoding a puromycin N-acetyl-transferase isolated from a ... Customers should verify that the version of a gene sold by InvivoGen is suitable for the customer needs. ... Synthetic Puromycin resistance gene in cloning plasmid. Resistance to the antibiotic puromycin is conferred by the Pac gene ...
C-E) Bioinformatics analysis of differentially regulated Tug1 target genes. (C and D) Biological processes GO terms from genes ... Zoom inset highlights the peak height, and the location of MACS verified the peak. Aligned reads were used for peak calling of ... Puromycin-resistant colonies were picked and analyzed for Tug1 or Ppargc1a expression levels by quantitative real time-PCR (qRT ... Tug1 mediates expression of PGC-1α pathway genes. (A) Gene expression analysis of RNA from pGIPZ-shControl (shCtrl) or shTug1 ...
Each expression pattern gene cluster from A is represented as a node connected to some of the top-ranked genes of each cluster ... Homologous and conditional miR-21 sequence was amplified from CJ7 ES cell-derived genomic DNA by PCR and verified by DNA ... The linearized targeting vector was electroporated into CJ7 ES cells, and PCR-mediated screening of 114 puromycin-resistant ... ToppGene Suite for gene list enrichment analysis and candidate gene prioritization. Nucleic Acids Res. 37(Web Server issue): ...
... and β-catenin genes (sh-β-catenin; with puromycin resistance), as well as their negative control (NC) nucleotide sequences, ... The sequences with high score were selected and the knockdown efficiency was verified by real-time PCR (RT-PCR) and ... After 7 days of G418 and/or puromycin treatment, cells were trypsinised and grown in a 96-well dish and expanded. shRNA ... To establish stable expressing cell lines, 400 μg/ml G418 and/or puromycin (Life Technologies) was added to the culture medium ...
B6.XCR1-lacZ mice (The Jackson Laboratory) is a strain in which the XCR1 gene has been replaced by the β-Gal reporter gene; ... the identity of the protein was verified by mass spectrometry. XCL1-StrepTag protein was purified from the supernatant using ... allows selection of positive transfectants by puromycin. Clones from limiting dilution cultures of transfected SL-3 cells were ... A BAC containing the human XCR1 gene was obtained from Childrens Hospital Oakland Research Institute (clone ID: RP11-527M10). ...
Gene Validation, LentiORF, MISSION® TRC3 Human LentiORF, MISSION® TRC3 ORF, Open Reading Frame, Protein Expression, Puromycin ... Verify the success of your overexpression experiments with BFP or GFP Expression. ... Synonym: Blasticidin ORF, Entry ORF, Gateway Cloning, Gene Expression, Gene Overexpression, Gene Validation, LentiORF, MISSION® ... Gene family sets are fully customizable allowing for complete flexibility to add or delete clone or gene content. ...
... and sequence-verified. Myc was expressed using murine stem cell virus (MSCV) retroviral vectors (11). For double-cooperativity ... design shRNAs were PCR-amplified from 97-mer oligonucleotides and cloned into MSCV-miR30-SV40-GFP or MSCV-miR30-PGK-Puromycin- ... coattenuation of the 8p22 gene Fgl1, the 8p21 gene Trim35, or the 8p23 gene Fbxo25 with Dlc1 by cotransduction of shRNA vectors ... A and B) Survival curves comparing high and low expression of indicated single genes (A) and combinations of two genes (B). (C ...
... is to create a shRNA library to enable the scientific community to use RNAi to determine the function of human and mouse genes ... Each hairpin sequence was cloned into the lentiviral vector (pLKO.1) and sequence verified. Multiple constructs (4 to 5) were ... Stable selection is also possible using the puromycin selectable marker. The pLKO.1 reagents can be packaged into virus ... Prospects for the therapeutic application of lentivirus-based gene therapy to HIV-1 infection. Curr. Gene Ther. 8(1), 1-8 (2008 ...
SelT and scr shRNAs were cloned into the lentiviral vector pVIRHD/EP containing a puromycin resistance gene. Constructs were ... Nucleotide sequence of all constructs was verified by DNA sequencing. ... Indeed, the UPR sensor gene Atf6 and the effector genes ChoP and Atf4 were significantly activated in shSelT‐treated cells. ... Bonkobara M, Das A, Takao J, Cruz PD, Ariizumi K (2003) Identification of novel genes for secreted and membrane‐anchored ...
... which contains the puromycin resistance gene (puro). For expression in Cos-1 cells, cDNAs encoding DNAM-1 or various PTKs were ... The resulting cDNAs were verified by sequencing (unpublished data). For expression in YT-S cells, cDNAs coding for DNAM-1 were ... Cells expressing the puromycin resistance marker (puro) or GFP alone were used as control. Isotype control is shown as filled ... Cells expressing the puromycin resistant marker (puro) alone are included as control. Isotype controls are shown as filled ...
Clones were screened with PCR, then verified by Sanger sequencing. Re-expression of Alg8, Ganab, and Sec61b cDNA was done with ... a large deletion was made between the sites of 2 guide RNAs in independent modified-pGL3-U6-sgRNA-PGK-puromycin (Addgene 51133 ... Gene-by-gene comparison showed enrichment of mutations for each of our candidate genes in PCLD cases: PKHD1, P = 2.34 × 10-8; ... In vitro gene knockout. All in vitro gene knockout studies were done in an immortalized mouse renal proximal tubule epithelial ...
The vector also contains a puromycin-resistance gene expression cassette. Puromycin-resistant clones were first selected to ... tCD4 gene expression unit. Recombinant antibody production of the selected clones was verified using an ELISA assay. ... The vector also contains a puromycin-resistance gene expression cassette. Puromycin-resistant clones are first selected to ... one for the neomycin resistance gene and one for the murine dhfr gene). All genes are under control of the promoter of the ...
To verify selective uptake by EpCAM+ tumor cells, we first used confocal fluorescence microscopy to compare internalization of ... using puromycin. The authors have not authenticated the cell lines used. Cell lines were obtained in 2012 and were expanded ... Gene knockdown. For in vitro gene silencing experiments, cells were used immediately after plating at 10,000 cells per well in ... To test RNA delivery, gene knockdown, and antitumor effects, siRNAs were incorporated to knock down a marker gene (eGFP), a ...
To verify that stable overexpression of VEGF-C was maintained in tumors during the experiment, Northern blot analysis was ... On the basis of the nucleotide sequence of the murine VEGF-C gene (DDB/EMBL/GenBank accession no. U73620), VEGF-C forward (5′- ... Two days after plating, 2 μg/ml puromycine were added for selection. Resistant colonies were picked 2 weeks after ... Subsequently, a 0.275-kV pulse with capacitance of 975 μF was applied using a Bio-Rad Gene Pulser II. After a 5-min incubation ...
Specific detection of dTIS11 by Western blotting was verified upon RNAi inhibition of the corresponding gene (Fig. 1C). Figure ... Puromycin was used at 50 μg/ml.. For pulse-chase analyses, S2 cells maintained in serum-free medium (ESF 921; Expression ... We thus verified dTIS11KRV5/HIS functionality in promoting ARE mRNA decay. To do this, we compared the abilities of dTIS11V5/ ... P-body formation is a consequence, not the cause, of RNA-mediated gene silencing. Mol. Cell. Biol. 27:3970-3981. doi:10.1128/ ...
... are ready to use gene editing systems designed to knockout or upregulate gene expression of human VPS52 ... gene to visually verify transfection. *The puromycin resistance and RFP genes are flanked by two LoxP sites to allow for ... Gene Info. Species. Gene Name. Gene ID. Chromosome Location. Isoform (mRNA) Accession #. Protein Accession #. OMIM™ Number. ... One plasmid in the pair contains a puromycin-resistance gene for selection; the other plasmid in the pair contains a GFP marker ...
... knockdown or upregulate mouse IFN-γRα gene expression ... gene to visually verify transfection. *The puromycin resistance ... Gene Info. Species. Gene Name. Gene ID. Chromosome Location. Isoform (mRNA) Accession #. Protein Accession #. OMIM™ Number. ... One plasmid in the pair contains a puromycin-resistance gene for selection; the other plasmid in the pair contains a GFP marker ... Each HDR Plasmid inserts a puromycin resistance gene to enable selection of stable knockout (KO) cells and an RFP (Red ...
Virostatic EBV genes have frequent variations in cancer and affect expression of host genes. A, Bar heatmaps showing EBV genes ... To better understand and verify the effect of IFN response pathways and EBV reactivation on PD-L1 and IDO1 expression in EBV+ ... The efficiency of EBNA1 knockout was validated by Western blotting at 3 days after puromycin selection. After knockout EBNA1 ... B, GSEA plot for EBV genes associated with 12 identified protein-coding virostatic genes, comparing EBV genes with frequent or ...
The successful replacement of both alleles by a puromycin and blasticidin resistance gene, respectively, was verified by PCR ... To replace the px I and II genes in the insect stage, procyclic T. brucei 449 cells were transfected with the vectors pHD1747- ... These may be main reasons why BS parasites in which the mitochondrial px III gene has been knocked out do not display any ... This allowed us to clone cell lines that lack the individual genes [19]. The selective knockout of the mitochondrial px III ...
To increase the utility of GigaMUGA for verifying the integrity of genetically engineered mice, a set of 87 probes was carried ... puromycin, hygromycin, and ampicillin. Probe sequences were designed against 51 bp of known construct sequence; alternate ... D) Genes in the complement pathway (black) and other genes (gray), from Ensembl. (E) Segmental duplications (black, from UCSC ... Probes targeted to complement-pathway genes detect copy-number variation. (A) Cluster plots for a probe in the C1s1 gene. ...
In podocytes, cAMP signaling activates cAMP response element-binding protein (CREB) to enhance expression of the gene encoding ... Proteomic analysis of glomeruli isolated from rats with puromycin-induced kidney disease and control rats indicated that ... We constructed and experimentally verified a β-adrenergic receptor-driven network with multiple feedback and feedforward motifs ... To determine how the motifs interacted to regulate gene expression, we mapped multicompartment dynamical models, including ...
  • To address these needs, we employed a transfer learning approach to derive gene-expression signatures from The Cancer Gene Atlas datasets that predict Fbw7 mutational status across tumor types and identified the pathways enriched within these signatures. (pnas.org)
  • Studies in isogenic colorectal cancer cell lines that differed in Fbw7 mutational status confirmed that Fbw7 mutations increase mitochondrial gene expression. (pnas.org)
  • Its expression confers puromycin resistance to pac-transfected cells. (invivogen.com)
  • Increasing evidence has identified that low expression of tumour suppressor genes and/or hyper-activation of oncogenes obviously accelerates the occurrence and development of lung SCC [ 4-7 ]. (portlandpress.com)
  • Comprehensive - Trusted TRC3 (The RNAi Consortium) content with ORF gene expression spanning 14,000+ human genes. (sigmaaldrich.com)
  • Verify the success of your overexpression experiments with BFP or GFP Expression. (sigmaaldrich.com)
  • Our ready-to-use MISSION® TRC3 Open Reading Frame (ORF) clones allow for stable integration, enrichment of cells, and long-term gene expression in difficult-to-transfect cell lines utilizing our best. (sigmaaldrich.com)
  • Here, we show that aptamer-siRNA chimeras (AsiC, an EpCAM aptamer linked to an siRNA sense strand and annealed to the siRNA antisense strand) are selectively taken up and knock down gene expression in EpCAM + cancer cells in vitro and in human cancer biopsy tissues. (aacrjournals.org)
  • Posttranscriptional regulation plays a central role in the control of gene expression in eukaryotic cells. (asm.org)
  • Functional analysis of the previously uncharacterized RAI2 protein revealed molecular interaction with CtBP transcriptional regulators and an overlapping function in controlling the expression of a number of key target genes involved in breast cancer. (aacrjournals.org)
  • Treatment of ABC lines with these inhibitors rapidly induced a series of gene expression changes that were attributable to cessation of constitutive IKK activity, similar to changes induced by acute expression of genetic inhibitors of NF-κB, confirming the effectiveness and specificity of this compound. (aacrjournals.org)
  • In the presence of tamoxifen, p65-ERD reversed the toxicity of IKK inhibition and restored expression of many NF-κB target genes. (aacrjournals.org)
  • Another subgroup of DLBCL, primary mediastinal B-cell lymphoma (PMBL), also expresses NF-κB target genes, and treatment of a PMBL cell line with an IKK inhibitor was toxic and induced gene expression changes of a distinct group of NF-κB target genes. (aacrjournals.org)
  • Diffuse large B-cell lymphoma (DLBCL), an aggressive non-Hodgkin's lymphoma, has been shown to consist of at least three molecular subgroups that differ in the expression of several hundred genes (1-5) . (aacrjournals.org)
  • The germinal center B cell-like (GCB) subgroup shares a broad gene expression program with normal germinal center B cells, which are therefore the presumed cell of origin for this subgroup (1, 3-5) . (aacrjournals.org)
  • The activated B cell-like (ABC) subgroup has lost expression of most germinal center B-cell genes and instead has gained expression of some plasma cell-associated genes, suggesting that it may be derived from a cell that is developmentally between the germinal center B cell and the plasma cell (1, 3-5) . (aacrjournals.org)
  • In adipocyte Senp2-deficiency mice, accumulation of the SUMOylated Setdb1 suppressed the expression of Pparg and Cebpa genes as well as lipid metabolism-related target genes, which would decrease the ability of lipid storage in adipocytes. (deepdyve.com)
  • The pLL-CMV-GFP-T2A-Puro Lenti-Labeler™ construct expresses copepod GFP from the CMV promoter, which delivers strong expression in most commonly-used cell lines (HeLa, HEK293, HT-1080, etc.), and coexpresses the puromycin resistance gene for selection in vitro prior to in vivo use. (bioscience.co.uk)
  • We now demonstrate that colonic tissue from Ier3 −/− mice subject of dextran sodium sulfate colitis exhibit greater Nrf2 activity than Ier3 +/+ mice, manifesting as increased nuclear Nrf2 protein level and Nrf2 target gene expression. (pubmedcentralcanada.ca)
  • 4 Acting mainly as a key regulator of the cellular response to oxidative and metabolic stress ( 5 ), Nrf2 induces the expression of a great number of antioxidative and phase II enzymes as well as a number of genes involved in cell growth and survival ( 6 ). (pubmedcentralcanada.ca)
  • The underlying metabolic shift is regulated at various levels including cell-signaling cascades or altered gene expression and protein degradation ( 1 , 3 ). (mcponline.org)
  • Most cell biology experiments utilize transient transfection protocols that afford peak gene expression between 24-96 hours post transfection. (mirusbio.com)
  • However, if sustained gene expression is required, generation of stable cell lines is a viable option. (mirusbio.com)
  • In such cases, a plasmid mixture containing 5 to 10 parts gene expression plasmid and 1 part antibiotic selection marker plasmid can be introduced into cells. (mirusbio.com)
  • After the stable cell line is generated, it is preferable to verify stable expression from a few cell passages before freezing stocks. (mirusbio.com)
  • However, the underlying transcriptional mechanisms that control the spatial and temporal expression of these genes are largely unknown. (biologists.org)
  • The pLL-EF1a-rFLuc-T2A-mRFP-mPGK-Puro Lenti-Labeler construct co-expresses rFLuc and mRFP from the EF1α promoter, which delivers moderate expression in most cell types (including primary and stem cells), and co-expresses the puromycin resistance gene for selection in vitro prior to in vivo use. (systembio.com)
  • The AHR/ARNT dimer regulates expression of a battery of genes involved in metabolism and elimination, including xenobiotic metabolizing enzymes CYP1A1 and CYP1B1 [ 10 , 11 ]. (hindawi.com)
  • Thererore, the "Rosa26" locus has since been used as a transgene insertion site that causes no apparent adverse effects on fitness, and permits stable gene expression. (genecopoeia.com)
  • To identify the target genes regulated by EGFR activation, we performed expression array analysis and found that the most commonly altered expression was from a gene module normally associated with IFN stimulation and Stat function. (rupress.org)
  • Classical NF-κB, a heterodimer composed of p50 and p65 subunits, is a potent activator of gene expression from NF-κB sites due to the presence of transactivating domains located in the C-terminal 120 amino acids of the p65 (also called RelA) protein ( 1 , 21 ). (asm.org)
  • GSK3 activity was decreased in mouse embryonic fibroblasts (MEFs) genetically and pharmacologically, and changes in the cyclical expression of core clock genes -mPer2in particular - were examined. (jcircadianrhythms.com)
  • Gene expression analysis showed that ALKBH3 does not directly influence the transcription of its target genes, but its depletion induces an upregulation of ALKBH3 non-bound inflammatory genes. (beds.ac.uk)
  • Cell surface proteins such as integrins have been suggested to transmit physical stimuli from the tendon ECM to regulate intracellular signaling pathways and gene expression 2 . (nature.com)
  • Western blot and real-time PCR were used to detect the expression levels of genes. (springer.com)
  • The fluorescent marker, puromycin resistance gene and shRNA are expressed from a single transcript making fluorescence a convenient and direct indicator of shRNA expression. (biocat.com)
  • PATIENTS AND METHODS: mRNA expression levels of 101 glycosyltransferase genes in TNBC patients were compared for correlation with patient survival using The Cancer Genome Atlas data. (bvsalud.org)
  • This approach uses a modified piggyBac transposon to generate libraries of mutagenized cells, each containing transposon insertions that randomly activate nearby gene expression. (biomedcentral.com)
  • Analysis of co-occurring transposons integration sites in single cell clone allows for the identification of genes that might act cooperatively to produce drug resistance a level of information not accessible using RNAi or ORF expression screening approaches. (biomedcentral.com)
  • Mutagenesis with mobile genetic elements that insert into the genome offers a great scope for screening as these provide readily detected tags to identify insertion sites, and can potentially either activate or disrupt gene expression. (biomedcentral.com)
  • The human WJ-MSCs were isolated and subsequently transduced with apelin-expressing lentiviral particles (WJMSCs-apelin), and expression was verified by flow cytometry, Western blot, ELISA, and RT-PCR analysis. (biomedcentral.com)
  • Therefore, we applied viral vector gene delivery technologies to ensure a constant bioactive apelin expression and secretion. (biomedcentral.com)
  • RBM38, a member of the RNA recognition motif (RRM) family of RNA binding proteins (RBPs), plays a fundamental role in the posttranscriptional control of gene expression and regulatory functions in human tumors. (biomedcentral.com)
  • Methods and compositions for reducing expression of genes on Chromosome 21 ("Chr 21") by targeting an XIST transgene to the Dual specificity tyrosine-phosphorylation-regulated kinase 1A (DYRK1A) gene or a Regulator of calcineurin 1 (RCAN1) gene, and cells and transgenic animals comprising an XIST transgene inserted into a DYRK1A or RCAN1 allele, e.g., cells and animals trisomic for human Chr 21 and mouse Chr 16. (patentsencyclopedia.com)
  • F) Potential CDX2-direct target genes based on ChIP-Seq and the alteration of expression by Cdx2-overexpression. (omicsdi.org)
  • G) Identification of CDX2 target genes by combining information on binding sites with gene expression response to Cdx2 over-expression Overall design: Chromatin IP against CDX2-Flag fusion protein. (omicsdi.org)
  • Open in a separate window Figure 2 Establishment of cell lines with stable HMGCS2 overexpression and knockdown: (A,B) Western blotting and QPCR were used to assess HMGCS2 gene expression in shHMGCS2 and HMGCS2-overexpressing cells. (bmn-673.com)
  • Genes and Biological Functions Affected by Downregulated Expression of HMGCS2 in HCC Cells To analyze alterations in the gene expression profile, total RNA was extracted from Huh-7 shlacZ and shHMGCS2 Xanthohumol cells for microarray experiments. (bmn-673.com)
  • In addition, the expression of genes related to ERK/MAPK signaling, PI3K/AKT signaling, EMT pathway, and molecular mechanism of cancer pathways were also identified by using IPA (Supplementary Figures S6CS9). (bmn-673.com)
  • Dharmacon is a distributor of multiple gene expression clone collections (cDNAs and ORFs). (horizondiscovery.com)
  • Here, we report that in T cell acute lymphoblastic leukemia (T-ALL) the MYC oncogene controls the expression of TET1 and TET2 to maintain 5-methylcytosine (5mC) and 5-hydroxymethylcytosine (5hmC) patterns, which is associated with tumor cell-specific gene expression. (biomedcentral.com)
  • Based on qPCR, expression of BCRP mRNA was knocked down in five clones with a maximum of 97% silencing in clone D. Silencing of BCRP gene expression was maintained for at least 25 passages. (aspetjournals.org)
  • In humans, high BCRP gene expression is linked to chemotherapeutic drug resistance. (aspetjournals.org)
  • In this study, The Cancer Genome Atlas (TCGA) dataset analysis revealed that upregulation of TSPAN12 gene expression was significantly correlated with patient survival, suggesting that TSPAN12 might be a potential prognostic marker for OC. (inforang.com)
  • This highlighted key pathways such as apoptosis and ubiquitination in GRN deficient human neurons, while revealing an unexpected major role for the Wnt signaling pathway, which was confirmed by analysis of gene expression data from postmortem FTD brain. (nih.gov)
  • Expression of hairpin was verified by visualization of robust RFP expression. (nih.gov)
  • C. Differential expression analysis was carried out and we observed hundreds of genes changing. (nih.gov)
  • Suppression of gene expression may be sufficient to limit the CAPN5 disease allele physiological activity and delay disease onset and progression. (biomedcentral.com)
  • Previous meta-analysis of gene expression profiles of 587 TNBC cases from 21 studies demonstrated high expression of Wnt signaling pathway-associated genes in basal-like 2 and mesenchymal subtypes of TNBC. (biomedcentral.com)
  • Hepatocellular carcinoma in these mice displays a cancer stem cell gene signature specified by the co-expression of ductal progenitor markers and oncofetal genes. (omicsdi.org)
  • RIN1 induced changes in gene expression that resembled siRNA silencing of RBPJ rather than inhibition at the level of NOTCH itself. (stanford.edu)
  • Most extensive coverage- Pre-cloned genome-wide arrayed libraries contain 50,000+ clone LentiORF library spanning 14,000+ human genes. (sigmaaldrich.com)
  • On the other hand, by combining shRNA with sgRNA, simultaneous gene silencing and genome targeting can be achieved. (frontiersin.org)
  • Murine studies have linked TGF-β signaling to emphysema, and human genome-wide association studies (GWAS) studies of lung function and COPD have identified associated regions near genes in the TGF-β superfamily. (elifesciences.org)
  • The kits are designed to specifically transfer your gene of interest, selection marker or other genetic element into the ROSA26 safe harbor site in mouse genome via TALEN-mediated homologous recombination. (genecopoeia.com)
  • The Genome-TALER™ mouse ROSA26 safe harbor gene knock-in kit includes one pair of ROSA26 TALENs, one donor cloning vector or one donor clone*, one positive control clone and one knock-in PCR verification kit. (genecopoeia.com)
  • One way to approach this is to build a minimal genome that includes only the genes essential for life. (sciencemag.org)
  • used a design, build, and test cycle to reduce this genome to 531 kb (473 genes). (sciencemag.org)
  • In 1995, we reported the first complete cellular genome sequences ( Haemophilus influenza , 1815 genes, and Mycoplasma genitalium , 525 genes). (sciencemag.org)
  • Comparison of these sequences revealed a conserved core of about 250 essential genes, much smaller than either genome. (sciencemag.org)
  • In 1999, we introduced the method of global transposon mutagenesis and experimentally demonstrated that M. genitalium contains many genes that are nonessential for growth in the laboratory, even though it has the smallest genome known for an autonomously replicating cell found in nature. (sciencemag.org)
  • hMPV is closely related to the important respiratory pathogen human respiratory syncytial virus (hRSV), which is also classified in the Pneumovirinae subfamily, but the order of the genes is different for the two viruses, and two nonstructural genes (NS1 and NS2) present in the RSV genome are missing from the genome of hMPV ( 50 ). (asm.org)
  • Genome-wide screens in Drosophila cells have offered numerous insights into gene function, yet a major limitation has been the inability to stably deliver large multiplexed DNA libraries to cultured cells allowing barcoded pooled screens. (elifesciences.org)
  • Effective therapeutic strategies for in vivo siRNA delivery to knockdown genes in cells outside the liver are needed to harness RNA interference for treating cancer. (aacrjournals.org)
  • Recent early-phase clinical trials have shown vigorous (75%-95%), sustained (lasting up to several months) and safe knockdown of a handful of gene targets in the liver using lipid nanoparticle-encapsulated or GalNAc-conjugated siRNAs ( 2-5 ). (aacrjournals.org)
  • The major obstacle to harnessing RNAi for treating most diseases, however, has yet to be solved, namely efficient delivery of small RNAs and gene knockdown in cells beyond the liver. (aacrjournals.org)
  • Here we develop a flexible, targeted platform for gene knockdown and treatment of basal-like TNBCs that might also be suitable for therapy against most of the common (epithelial) cancers. (aacrjournals.org)
  • This allows one to examine the effect of loss of gene function over a large series of gene knockdown efficiencies. (sigmaaldrich.com)
  • Establishment of Cell Lines with Steady HMGCS2 Overexpression and Knockdown To clarify the tasks of HMGCS2 Xanthohumol manifestation in liver tumor cells, the HMGCS2 gene was either knocked down or overexpressed in Huh-7 and Hep3B cells by lentivirus infection. (bmn-673.com)
  • Functional ketogenesis activity in both HMGCS2 knockdown and HMGCS2-overexpressing cells was verified by a colorimetric HB assay (Figure 2C). (bmn-673.com)
  • To reveal the pathways dysregulated by HMGCS2 gene knockdown, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis was performed. (bmn-673.com)
  • Cellular movement, cell-to-cell signaling and interaction, and cell death and survival were markedly changed after Xanthohumol HMGCS2 gene knockdown (Figure 3C). (bmn-673.com)
  • Most genes (152/153) common to both hairpins are differentially expressed in the same direction relative to controls, further confirming that this list of genes is robustly related to GRN knockdown. (nih.gov)
  • Lentivirus-mediated shRNA knockdown of STMN1 gene manifestation shSTMN1 and non-silencing GV248 control vector were purchased from Gene Chem Co. (Shanghai, China). (lecollege.org)
  • Genes involved in mitochondrial function were highly enriched in pan-cancer signatures that predict Fbw7 mutations. (pnas.org)
  • It has been documented that more than 50% human tumors bear p53 gene mutations, of which missense mutations account for over 70% of all p53 mutants 2 . (nature.com)
  • As an example, if someone has genetic mutations that alter the activity of a gene called TGFB2 , their risk of developing emphysema increases. (elifesciences.org)
  • This revealed that certain mutations near the TGFB2 gene were more common in patients with emphysema. (elifesciences.org)
  • Mutations of IDH genes result in production of the oncometabolite 2-hydroxyglutarate (2-HG) instead of α-ketoglutarate [ 6 ]. (aging-us.com)
  • We further found that SET domain bifurcated 1 (Setdb1) was a SUMOylated protein and that SUMOylation promoted Setdb1 occupancy on the promoter locus of Pparg and Cebpa genes to suppress their expressions by H3K9me3. (deepdyve.com)
  • This effect was independent of age at diagnosis, treatment received and hypermethylation of the O(6)-methylguanine methyltransferase (MGMT) DNA repair gene promoter. (biomedsearch.com)
  • Target genes were indicated in (G). Primers flanking a promoter region of Hbb-b1 and Pou5f1 as well as a gene desert region in chromosome 3 were used as negative controls. (omicsdi.org)
  • Primers flanking of Actb gene promoter were used for normalization. (omicsdi.org)
  • Smyd3 specifically interacts with H3K4Me3-modified histone tails and is recruited to the core promoter regions of many but not all active genes. (omicsdi.org)
  • Validate your gene(s) of interest from your RNAi or CRISPR experiments. (sigmaaldrich.com)
  • Target-specific CRISPR Plasmids for both gene knockout and activation are available. (scbt.com)
  • The CRISPR Cas9 nuclease, in combination with single guide RNAs (sgRNAs), is widely used to create targeted genomic modifications, from gene knockouts, mutagenesis, fusion tagging, and more, in eukaryotic cells and animal models. (genecopoeia.com)
  • Under basal growth conditions, 1235 genes were essential for cell fitness at a false-discovery rate of 5%, representing the highest-resolution fitness gene set yet assembled for Drosophila , including 407 genes which likely duplicated along the vertebrate lineage and whose orthologs were underrepresented in human CRISPR screens. (elifesciences.org)
  • Scientists often use a technique called CRISPR to edit genes. (elifesciences.org)
  • Choose from 3 different gateway-adapted vectors: Entry, Blasticidin, or Puromycin. (sigmaaldrich.com)
  • The monocytic cell line THP-1 and the retina pigmented ephitelium like cell line ARPE-19 were transduced with lentiviral vectors to generate stable cell lines by selecting for puromycin resistant cells. (arvojournals.org)
  • The Lenti-Labeler™ vectors from System Biosciences provide an easy way to label cells in vitro for downstream in vivo applications.The Lenti-Labeler™ comes with a range of selection markers (Puro and Blasticidin), reporter genes (GFP, RFP, BFP and luciferase) and two different promoters (CMV and EF1a). (bioscience.co.uk)
  • With their sequences verified, all of the Lenti-Labeler™ vectors are based on time-tested and proven third generation backbones to deliver high quality and performance for nearly all potential cell labelling applications. (bioscience.co.uk)
  • An adenoviral strong splice acceptor, typically used in gene trap vectors, is fused upstream of the his3 stuffer fragment to prevent splicing around the stopper (in the case that transcription isn't completely silenced). (jax.org)
  • To overcome this bottleneck, we developed an unbiased genetic approach to identify new genes and pathways that mediate stress resistance and longevity in the mouse. (frontiersin.org)
  • Among the genes and pathways controlled by TET1, we found ribosomal biogenesis and translational control of protein synthesis highly enriched. (biomedcentral.com)
  • To identify the pathways downstream of GRN, we used weighted gene coexpression network analysis (WGCNA) to develop a systems-level view of transcriptional alterations in a human neural progenitor model of GRN-deficiency. (nih.gov)
  • In these organs Smyd3 is functioning in the nucleus as a direct transcriptional activator of several key genes involved in cell proliferation, epithelial-mesenchymal transition, JAK/Stat3 oncogenic pathways, as well as of the c-myc and b-catenin oncogenes. (omicsdi.org)
  • The nonsegmented, negative-strand RNA genomes of the MPVs are approximately 13.3 kb long and contain eight genes that are separated by gene start (GS) and gene end (GE) sequences and encode nine putative proteins: nucleoprotein (N), phosphoprotein (P), matrix (M) protein, fusion (F) protein, small hydrophobic surface protein (SH), major attachment glycoprotein (G), major polymerase subunit (L), transcription elongation factor (M2.1), and RNA synthesis regulatory factor (M2.2). (asm.org)
  • It enables researchers to modify DNA sequences to selectively alter the purpose of a gene or even turn it off to find out what it does. (elifesciences.org)
  • RNA-based gene silencing methods generate gene-specific complementary RNA sequences that form double stranded RNA within cells. (biomedcentral.com)
  • All of the gene family sets can be used at the benchtop for focused pathway analysis and building your disease model of interest. (sigmaaldrich.com)
  • Different gene pathway of interest? (sigmaaldrich.com)
  • The NF-κB pathway also influences proliferation, which may involve NF-κB target genes such as cyclin D2 , c- myc , and IL-6 (18-20) . (aacrjournals.org)
  • Thus, the novel IGF2.sgRNA-LIG4.shRNA-IGF2.sgRNA structure was constructed to enhance the sgRNA/Cas9-mediated HDR-based IGF2 SNP editing by silencing the LIG4 gene, which is a key molecule of the HDR's competitive NHEJ pathway. (frontiersin.org)
  • The NHEJ pathway generates stochastic nucleotide insertions and deletions (Indels) at the target locus resulting in open reading frame (ORF) shift and loss-of-function of target genes. (frontiersin.org)
  • Tissue and cellular imaging confirmed that SANPCs contained key elements of glutamatergic neurotransmitter system, expressing genes encoding glutamate synthesis pathway (Gls), ionotropic and metabotropic glutamate receptors (Grina, Gria3, Grm1 and Grm5), and glutamate transporters (Slc17a7). (bvsalud.org)
  • This strategy offers an efficient way to select for new mutants expressing a stress resistance phenotype, to rapidly identify the causative genes, and to develop mice for in vivo studies. (frontiersin.org)
  • Members of several gene families are important for blastocyst formation, including claudins, zonal occludins (TJ proteins), occludins, jam proteins, cell polarity proteins, Na/K-ATPases and aquaporins. (biologists.org)
  • These interactions physically link the AHR to proteins involved in the immune and cellular stress responses, gene regulation not mediated directly via the traditional AHR:ARNT heterodimer, and mitochondrial function. (hindawi.com)
  • The DNA sequence of these genes were modified by optimizing the codon usage, reducing or eliminating the CpG motifs and avoiding secondary DNA structures without changing the amino acid sequence of the wild type proteins. (docplayer.net)
  • The chimeric proteins that are encoded from the fusion genes in the leukemic cells could in basic principle serve as tumor markers but there is a lack of appropriate antibodies for the development of immunoassays (7). (himafund.org)
  • The last two proteins (collectively named M2 here) are thought to be expressed from overlapping open reading frames in the M2 gene ( 16 , 83 ). (asm.org)
  • HMGCS2 proteins and mRNA manifestation in the Hep3B and Huh-7 cell lines had been verified by traditional western blotting and quantitative real-time polymerase string response (QPCR) assays, respectively (Shape 2A,B). Proteins quantification was performed utilizing the ImageJ program (Supplementary Shape S1). (bmn-673.com)
  • By targeting the mouse orthologs of genes frequently deleted on human 8p22 and adjacent regions, which are lost in approximately half of several other major epithelial cancers, we provide evidence suggesting that multiple genes on chromosome 8p can cooperatively inhibit tumorigenesis in mice, and that their cosuppression can synergistically promote tumor growth. (pnas.org)
  • Here we explore the hypothesis that chromosome 8p deletions arise owing to selection for the attenuation of multiple genes. (pnas.org)
  • The E. coli rnpb terminator allows efficient transcription termination of the Puro-lowCpG gene. (invivogen.com)
  • The resulting JCV-syn3.0 retains genes involved in key processes such as transcription and translation, but also contains 149 genes of unknown function. (sciencemag.org)
  • Intro Chromosomal translocations have a primary part in the pathogenesis of leukemia because they impact specific genes encoding transcription factors or additional cell-cycle regulators. (himafund.org)
  • Further, we propose that ALKBH3 is an intrinsic DNA repair protein that suppresses transcription associated DNA damage at highly expressed genes and thereby plays a role to maintain genomic integrity in ALKBH3-overexpressing cancer cells. (beds.ac.uk)
  • Bioinformatics, chromatin immunoprecipitation and transcriptional analysis revealed that Tcfap2c binds and regulates a diverse group of genes expressed during blastocyst formation. (biologists.org)
  • Although DLC1 is at an epicenter of deletions, these deletions are frequently much larger and reduce the dosages of tens or hundreds of genes, often encompassing the entire 8p22 cytoband and beyond ( 2 , 5 , 6 ). (pnas.org)
  • Some of the developmental genes involved in self-renewal maintenance, differentiation and regeneration are also proto-oncogenes such as My c. (biologists.org)
  • Despite recent identification of many oncogenes and tumor suppressor genes important for the development and treatment of NSCLC, overall 5-year survival remains poor at 15% ( 2-6 ). (aacrjournals.org)
  • Host cell transcriptional changes in response to EBV infection classified tumors into two molecular subtypes based on patterns of IFN signature genes and immune checkpoint markers, such as PD-L1 and IDO1. (aacrjournals.org)
  • Stable cell line generation is made possible by the use of positive selection markers such as G418, hygromycin B, puromycin resistance, etc. (mirusbio.com)
  • Moreover, the fusion genes that are created as a result of the translocations constitute important tumor markers whose detection and/or quantification assist in the analysis, prognosis, monitoring the response to treatment and detection of minimal residual disease (4C6). (himafund.org)
  • Sigma Bioinformatics has methodically mapped all TRC3 ORF clones to each current gene set. (sigmaaldrich.com)
  • Each gene family set is representationally titered (10% of clones). (sigmaaldrich.com)
  • Standard Smyd3-deficient (Smyd3-KO) mice were generated using gene-trap ES cell clones (AS0527 from International Gene Trap Consortium), in which a selection cassette, containing the splice acceptor site from mouse EN2 exon 2 followed by the beta-galactosidase and neomycin resistance gene fusion gene and the SV40 polyadenylation sequence was inserted into the 5th intron of the Smyd3 gene. (omicsdi.org)
  • One mechanism causing persistent Nrf2 activation is the adaptation of epithelial cells to oxidative stress during chronic inflammation, e.g. colonocytes in inflammatory bowel diseases, and the multifunctional stress response gene immediate early response-3 ( IER3 ) has a crucial role under these conditions. (pubmedcentralcanada.ca)
  • showed that, in healthy lung cells called fibroblasts, the stretch of DNA that was mutated in patients with emphysema touched the part of TGFB2 that controls when the gene is activated. (elifesciences.org)
  • Deleting that same stretch of DNA in the fibroblasts meant the cells could no longer activate the TGFB2 gene as efficiently. (elifesciences.org)
  • Often, researchers create many sgRNAs and deliver them to a large pool of cells with the help of viruses, so that each cell gets a different sgRNA that mutates a different gene. (elifesciences.org)
  • When the cells are then treated with a specific drug, the composition of the sgRNAs in the pool changes, depending on which genes are needed to withstand the drug, and which genes - when turned-off - create cells that are resistant to the drug. (elifesciences.org)
  • This allowed to identify over 1,000 genes necessary for cells to multiply properly, many of which had not been studied before. (elifesciences.org)
  • Puromycin (1 g/mL) was put into the culture moderate to choose the gene-transfected steady cells. (bmn-673.com)
  • There were no significant morphological changes between the different HMGCS2 gene-modified cells (Supplementary Figure Rabbit Polyclonal to ZP4 S2). (bmn-673.com)
  • SQ22,536 [9-(tetrahydrofuryl)-adenine] and 2′,5′-dideoxyadenosine (ddAd) are effective and potent AC inhibitors in HEK293 cells expressing a cAMP response element (CRE) reporter gene, and MDL-12,330A [cis-N-(2-phenylcyclopentyl)azacyclotridec-1-en-2-amine hydrochloride] is not. (aspetjournals.org)
  • While studying the effect of proteoglycan composition on nephritic permselectivity, it was noted that puromycin treatment of human glomerular endothelial cells (HGEC) altered the sulfation level of GAG chains on proteoglycans such as perlecan, which in turn caused a decrease in the stability of the GAG chains. (wikipedia.org)
  • In short, we achieved effective IGF2 SNP editing by using the combined sgRNA-shRNA/StCas9 strategy, which will facilitate the further production of base-edited animals and perhaps extend for the gene therapy for the base correction of some genetic diseases. (frontiersin.org)
  • However, it's always confronted with the concern of genetic safety, when creating gene knock-out/in animals. (frontiersin.org)
  • A better understanding of what genes do could help to identify potential genetic weaknesses in certain types of cancer or other diseases, which may lead to the development of more effective treatments. (elifesciences.org)
  • Forward genetic approaches using mobile genetic elements provide a powerful alternative method for gene discovery that can overcome many of the limitations of reverse genetic approaches. (biomedcentral.com)
  • In addition, in human HCC patients, the combined down-regulation of functionally validated 8p TSGs is associated with poor survival, in contrast to the down-regulation of any individual gene. (pnas.org)
  • Early studies on the tumor-suppressor genes (TSGs) RB and TP53 suggested that such deletions can arise as a single mechanism for loss of heterozygosity and, consequently, it is often assumed that they provide a "second-hit" event to inactivate a single TSG ( 1 ). (pnas.org)
  • Stable selection is also possible using the puromycin selectable marker. (horizondiscovery.com)
  • The cis approach is generally easier and has a higher likelihood of producing drug-resistant stable transfectants that express the gene of interest. (mirusbio.com)
  • The large chromosomal deletions frequently observed in cancer genomes are often thought to arise as a "two-hit" mechanism in the process of tumor-suppressor gene (TSG) inactivation. (pnas.org)
  • L1CAM is a novel prometastatic gene in NSCLC, and its downregulation may effectively suppress NSCLC tumor growth and metastasis. (aacrjournals.org)
  • Bloodstream Trypanosoma brucei in which the genes for cytosolic lipid hydroperoxide-detoxifying peroxidases have been knocked out undergo an extremely rapid membrane peroxidation and lyse within less than two hours when they are cultured without an exogenous antioxidant. (prolekare.cz)
  • Mechanistically, adipocyte Senp2 deficiency caused the downregulation of Pparg and Cebpa as well as their downstream target genes related to lipid storage. (deepdyve.com)
  • In cancer samples, hundreds of novel frequent missense and nonsense variations in virostatic genes were identified, and variant genes failed to regulate their viral and cellular targets in cancer. (aacrjournals.org)
  • We propose a model in which Tcfap2c acts in a hierarchy to facilitate blastocyst formation through transcriptional regulation of core genes involved in tight junction assembly, fluid accumulation and cellular proliferation. (biologists.org)
  • Our approach may be applied to study other complex cancer genes. (pnas.org)
  • However, genomic approaches have not conclusively identified a definitive TSG within some cancer-associated deletions, raising the possibility that they occur through genomic instability or selection for the reduced activity of multiple genes. (pnas.org)
  • The Lentiviral cassette contains three major functional elements, shRNA, a puromycin selection marker & a tGFP reporter. (origene.com)
  • The STOP element incorporates a PGK-puromycin selection cassette at the 5' end in an opposite directional orientation. (jax.org)
  • Positive controls can be used for measuring transduction efficiency and optimizing ORF delivery in cell lines that do not survive any puromycin exposure. (sigmaaldrich.com)
  • The CARD domain was amplified from the human apoptosis-associated speck-like protein containing a CARD (ASC) gene and fused to the cell-penetrating peptide sequence derived from the HIV Tat protein using PCR. (arvojournals.org)
  • RNA interference experiments demonstrated that Tcfap2c regulates genes important for tight junctions, cell polarity and fluid accumulation. (biologists.org)
  • A goal in biology is to understand the molecular and biological function of every gene in a cell. (sciencemag.org)
  • Using this approach we successfully identify genes involved in paclitaxel resistance in a variety of cancer cell lines, including the multidrug transporter ABCB1, a previously identified major paclitaxel resistance gene. (biomedcentral.com)
  • In this study, we report a novel approach to treat type 2 diabetic rats that combines apelin gene therapy with WJ-MSC cell therapy, which could provide a promising therapeutic option for management of type 2 diabetes clinically. (biomedcentral.com)
  • Ectopic MVP overexpression in the MVP-negative lung cancer cell model H65 led to a downregulation of three known IFN-γ-regulated genes, namely ICAM-1, CD13 and CD36. (biologists.org)
  • Nlrp1 genes from sensitive rat macrophages can confer sensitivity to this rapid cell death when expressed in Toxoplasma resistant rat macrophages. (prolekare.cz)
  • GV248 cloning vector contained eGFP and elements that were required for packaging of CK-1827452 inhibitor the manifestation create into virions, as well as the puromycin-resistant gene. (lecollege.org)
  • The SNP within intron 3 of the porcine IGF2 gene (G3072A) plays an important role for muscle growth and fat deposition in pigs. (frontiersin.org)
  • Human miR-17-92 is located within intron 3 of the C13orf25 gene at 13q31.3, a region frequently amplified in many hematopoietic and solid malignancies ( 6 ). (spandidos-publications.com)
  • Thus, for the functional characterization of emphysema GWAS loci, it is necessary to localize causal variants in regulatory elements and identify the gene(s) regulated by that element. (elifesciences.org)
  • UCSC Mouse Mm9 browser view of Hoxa7 gene locus after mapping CDX2 ChIP-Seq tags locations in the wiggle format. (omicsdi.org)
  • Furthermore, we observed that the Wnt receptor Fzd2 was one of only a few genes upregulated at 6 weeks in a GRN knockout mouse, and that FZD2 reduction caused increased apoptosis, while its upregulation promoted neuronal survival in vitro. (nih.gov)