Plasmid cho cells. Medical search. Frequent questions

This host-to-host transfer of genetic material is one mechanism of horizontal gene transfer, and plasmids are considered part of the mobilome. (wikipedia.org)
A typical bacterial replicon may consist of a number of elements, such as the gene for plasmid-specific replication initiation protein (Rep), repeating units called iterons, DnaA boxes, and an adjacent AT-rich region. (wikipedia.org)
Plasmids almost always carry at least one gene. (wikipedia.org)
CHO-K1 cells were transfected with human ER and SUMO1 expressing plasmids together with the CRH promoter reporter gene. (nel.edu)
Cell Line Development: Stable cell lines are widely used in a number of important applications including biologics (e.g. recombinant protein and monoclonal antibody) production, drug screening, and gene functional studies. (labroots.com)
We demonstrate that relatively short single-stranded oligodeoxynucleotides, 25-61 bases homologous to the target sequence except for a single mismatch to the targeted base, are capable of correcting a single point mutation (G to A) in the mutant beta-galactosidase gene, in nuclear extracts, episome, and chromosome of mammalian cells, with correction rates of approximately 0.05%, 1% and 0.1%, respectively. (nih.gov)
If CRISPR-Cas plasmids were to be used in humans, these plasmids could pose hazards by integrating a Cas gene into the genome. (idtdna.com)
CHO suspension clonal stable cell lines are generated by electroporation using proprietary mammalian expression vector system with multiple selectable markers to express single gene of interest. (welgen.com)
Despite multiple advances in cell engineering, the rapid creation of robust and multi-gene cell lines remains of reach - until now. (genelink.com)
A stable cell line is a population of cells that has been genetically modified to express a desired gene or protein in a stable and heritable manner. (altogenlabs.com)
The generation of a stable cell line typically involves introducing a transgene or gene of interest into the genome of a host cell and selecting for cells that have integrated the gene and are expressing it at a stable level over time. (altogenlabs.com)
Stable cell lines over-expressing a protein of interest (or shRNA construct targeting a gene of interest) are important research tools, particularly for the production of recombinant antibodies and other proteins, the study of gene functions, drug screening, and other applications. (altogenlabs.com)
All clonal stable cell lines are developed based on individual client specifications, using highly efficient gene delivery technologies to produce stably expressing clonal cell lines in just 28 days. (altogenlabs.com)
Typically, antibiotic resistance or fluorescent reporter gene markers are incorporated into the plasmid DNA construct to facilitate selection process. (altogenlabs.com)
Choose a host cell line: The first step in generating a stable cell line is to choose a suitable host cell line that can be efficiently transfected and stably maintain the introduced gene. (altogenlabs.com)
CP-PVY mRNA knockdown efficiency was analyzed by RT-PCR and real-time PCR of CHO-k cells co-transfected with a CP gene construct and siRNAs. (virosin.org)
Transmembrane and intracellular signal transduction mechanisms are the focus of our group, especially understanding how extracellular stimuli control cell growth and division, cell morphology, and gene expression at the biochemical level. (berkeley.edu)
Transient gene expression in mammalian cells provides reasonable amounts of recombinant proteins and antibodies which is sufficient for most applications. (invivo.de)
Purified plasmids for transfection along with the handle plasmid pEGFP-N2 (Clontech) have been prepared applying an EndoFree Plasmid Maxi Kit (Qiagen, Valencia, CA). For stable cell line generation all plasmids except p1.2-HygroeGFP had been linearized by restriction with PvuI by cutting inside the ampicillin resistance gene bla sequence. (dna-pkinhibitor.com)
The plasmid p1.2-HygroeGFP was restricted with BspHI, which introduced two breaks close to the bla gene.Cell cultureFragments corresponding towards the upstream and downstream flanking regions (8532?2603 and 14545?8794 sequences of [GenBank:AY188393]) of your CHO elongation element 1 gene have been obtained by PCR using CHO DG44 cell (Invitrogen) genomic DNA as a template. (dna-pkinhibitor.com)
Plasmids p1.2 were transfected by electroporation in Gene Pulser Electroporation Buffer (Bio-Rad, Hercules, CA) making use of a P2X1 Receptor Antagonist Molecular Weight cuvette using a four mm gap with 7.5 million cells and 15 g of linearized DNA for every transfection. (dna-pkinhibitor.com)
Electric field-mediated gene transfer in mammalian cells (electrotransformation) depends on the pulsing conditions (field intensity, pulse duration, number of pulses). (uni-bielefeld.de)
Pulsing conditions inducing reversible permeabilization of the cell plasma membrane are not sufficient to induce gene transfer. (uni-bielefeld.de)
Wolf H, Rols MP, Boldt E, Neumann E, Teissie J. Control by pulse parameters of electric field-mediated gene transfer in mammalian cells. (uni-bielefeld.de)
Control by pulse parameters of electric field-mediated gene transfer in mammalian cells", Biophysical Journal , vol. 66, 1994, pp. 524-531. (uni-bielefeld.de)
Compared with other eukaryotic expression systems, the expression of the target gene in mammalian cells is similar to that of the native protein in the type and manner of the glycosylation, and can be correctly assembled into the multi-subunit protein. (abvigen.com)

The spike (S) protein of SARS-CoV-2 mediates viral receptor binding and membrane fusion, both of which are essential for viral infection of host cells. (nature.com)
ACP-ADRβ2 fusion protein gives a cell surface localized signal when labeled with CoA substrates. (neb.com)
Each reagent was used to transfect HeLa cells in a 96-well format at the indicated doses with an emerald green fluorescent protein (GFP)-expressing vector. (thermofisher.com)
The company is currently evaluating the serum-free supplement on other aspects of bioprocessing, including single-use bioreactors, protein quality, and cell metabolism. (genengnews.com)
The earliest biochemical evidence on the existence of cell-free soluble LH receptor was the purification of an hCG-binding protein, relative molecular mass of 65K (M r , 65K) from porcine follicular fluid and was based on gel filtration followed by affinity chromatography [ 9 ]. (biomedcentral.com)
15 nm, we demonstrate that single-protein resolution can be achieved for biomolecules in whole intact cells. (nature.com)
2. Pilot small-scale transient transfection into HEK293 cells (Tuna293™ Process) or CHO cells (TunaCHO™ process) is performed, followed by the listed purification method to assess expression levels and protein yield. (curiaglobal.com)
Stable cell lines expressing recombinant proteins are generated by experienced scientists working on protein engineering, expression, and purification, and cell line development services for decades. (welgen.com)
Stable cell lines developed with our proprietary protein expression system with multiple selectable markers is very productive in expressing target proteins. (welgen.com)
Ribonucleoprotein (RNP) complexes composed of PPIB (cyclophilin B) targeting two part gRNA (IDT) and Cas9 protein (PNA Bio) were delivered into HEK293T/17 and U2OS cells using TransIT-X2® Dynamic Delivery System or Lipofectamine® CRISPRMAX™ or Lipofectamine® RNAiMAX or Lipofectamine® 3000 in a 24-well format according to the manufacturers' protocol. (bioscience.co.uk)
Altogen Labs offers several stable cell line generation services: protein overexpressing cell lines, RNAi knockdown cell lines, and reporter stable cell lines (luciferase, GFP, RFP, YFP). (altogenlabs.com)
The experiments showed that one end of the protein, called the N-terminus, is involved in the movement of electrical charges across the cell membrane and is able to cause cell degeneration. (elifesciences.org)
By contrast, the other end of the protein, the C-terminus, acts as a regulator for the N-terminus and can prevent cell degeneration. (elifesciences.org)
The Wee1 class of protein-tyrosine kinase has an important role in cell cycle control. (berkeley.edu)
We investigated control mechanisms that regulate the activity, localization, and stability of Wee1, especially the bud neck-localized protein kinase Hsl1 and its more distant paralogs (Gin4 and Kcc4), in particular their recruitment to septin filaments, which assemble at the presumptive site of cell division. (berkeley.edu)
We also studied the roles of other classes of protein kinases (Cla4) and additional post-translational modifications (SUMOylation) in septin complex assembly, formation of different septin-based supramolecular ensembles, disassembly of septin-containing structures, and the function of septin organization in the events required for cell division and membrane septation during cytokinesis. (berkeley.edu)
For clients with difficult-to-transfect cells or not enough time or resources on their hand, Synvolux also offers custom screening and optimization services to identify the most suitable transfection reagent from Synvolux' library of >100 distinct transfection reagents in combination with DNA, siRNA, mRNA, or protein/peptides. (synvoluxproducts.com)
Using these customized vectors, high protein yields and a substantially reduced timeline from transfection to stable CHO cell pools could be achieved. (invivo.de)
After detailed rounds of discussion with our scientific experts, we help you decide if a stable cell line needs to be developed or if we can produce the required protein yields from a stable cell pool. (invivo.de)
During stable transfection, the DNA of interest is integrated into the genome of the host cell to allow steady production of the target protein. (invivo.de)
This creates a pool of cells which all express the same protein or antibody, but with varying degrees of expression efficiency. (invivo.de)
Trinomab's process development platform consists of four parts: cell culture, protein purification, formulation prescription, and analytical science. (trinomab.com)
Expression of the fusion protein cloned in pSNAP-CaaX can be achieved by transiently transfecting cells in culture with standard transfection protocols. (neb.com)
The K V 10.1-specific scFv62 antibody -TRAIL fusion protein was expressed in CHO-K1 cells, purified by chromatography and tested for biological activity. (biomedcentral.com)
In co-cultures with K V 10.1-positive cancer cells the fusion protein also induced apoptosis in bystander K V 10.1-negative cancer cells, while normal prostate epithelial cells were not affected when present as bystander. (biomedcentral.com)
TRAIL is normally present as a membrane protein (memTRAIL) on immune effector cells, like natural killer cells. (biomedcentral.com)
Pili (or fimbriae), fibrils, and a protein called Hia mediate adherence of Hib to cells of the human respiratory tract. (medscape.com)

pACP-ADRβ2 has performed well in transient transfection of CHO-K1, COS-7, U-2 0S and NIH 3T3 cells. (neb.com)
11 ] first reported the production and secretion of soluble LH receptor following transient transfection of a naturally truncated variant of the receptor in COS cells. (biomedcentral.com)
A total of six siRNAs were designed and screened through transient transfection assay and knockdown in expression of CP-PVY mRNA was calculated in CHO-k cells. (virosin.org)
pSNAP-CaaX has performed well in stable and transient transfection of CHO-K1 and NIH 3T3 cells. (neb.com)

Commonly used host cell lines include Chinese hamster ovary (CHO), HEK293, and HeLa cells. (altogenlabs.com)

Artificial plasmids are widely used as vectors in molecular cloning, serving to drive the replication of recombinant DNA sequences within host organisms. (wikipedia.org)
A DNA fragment encoding eGFP and also a consensus Kozak sequence (GCCGCCATGG) [14] was obtained by PCR with adaptor primers and the pEGFP-C2 plasmid (Clontech, Mountain View, CA) as a template after which cloned in to the polylinker area of p1.1 and p1.2 vectors, thereby resulting in p1.1(EBVTR-)eGFP, p1.1eGFP, p1.2HygroeGFP, p1.2-NeoeGFP and p1.2-ZeoeGFP expression plasmids. (dna-pkinhibitor.com)

Lipofectamine 3000 Transfection Reagent delivers a high efficiency for hard-to-transfect cell lines. (thermofisher.com)
When performing transfections in hard-to-transfect cell lines, Lipofectamine 3000 reagent can be an ideal choice. (thermofisher.com)
It delivers exceptional transfection efficiency with improved cell viability for the widest range of hard-to-transfect and common cell types including HEK 293, HeLa, LNCaP, HepG2, and A549 cell lines (Figure 1) . (thermofisher.com)
Each reagent was used to transfect HEK 293, HeLa, LNCaP, HepG2, and A549 cell lines in a 96-well format, and GFP expression was analyzed 48 hours post-transfection. (thermofisher.com)
We were very happy and surprised to see Lipofectamine® 3000 reagent provide a more than 10-fold difference in transfection efficiency in our difficult-to-transfect cell line. (thermofisher.com)
4-8 µl TransIT-X2™ used to transfect 2 µg plasmid DNA encoding EGFP into A549, CHO-K1, HepG2, LNCaP, MDCK, PC12, primary human mammary epithelial cells (HMEC) and normal human dermal fibroblasts (NHDF). (bioscience.co.uk)
TransIT-X2 was used to transfect plasmid Cy®5 labeled DNA encoding nuclear YFP and Cy®3 labeled siRNA into HeLa cells. (bioscience.co.uk)
Transfect the cells: The next step is to introduce the vector containing the transgene into the host cells. (altogenlabs.com)

U2OS.EGFP cells caused by inactivating frameshift insertion/deletion (indel) mutations introduced by error prone non-homologous end-joining (NHEJ) repair of nuclease-induced double-stranded breaks (DSBs) (Fig. 1a and Methods ). (cdc.gov)

Back in the 1950's, cells from Henrietta Lacks (now called HeLa cells) were isolated and cultivated in horse serum," recalls Adam Elhofy, Ph.D., CSO, Essential Pharma. (genengnews.com)

Plasmids are transmitted from one bacterium to another (even of another species) mostly through conjugation. (wikipedia.org)

Lipofectamine 3000 reagent provided higher GFP transfection efficiency than Lipofectamine 2000 and FuGENE HD reagents for all five cell lines. (thermofisher.com)
This superior transfection performance allows you to reduce the reagent dose, thus helping lower toxicity risk when transfecting your cell line. (thermofisher.com)
Lipofectamine 3000 reagent is a specially formulated gentle transfection reagent that is optimized for plasmid delivery into CHO cells with high efficiency and low cell toxicity. (thermofisher.com)
The easy protocol, exceptional plasmid DNA delivery, and the ability to effectively generate stable cell lines makes Lipofectamine 3000 reagent, the perfect reagent for CHO cell transfection experiments. (thermofisher.com)
Further, in the presence of this biochemical reagent, we were able to reduce the required plasmid concentration by ∼70% without compromising the transfection efficiency. (elsevierpure.com)
For direct colony generation in 96-well culture plates, transfection was performed making use of Fugene HD reagent (Promega), containing 60 g of DNA and 180 l with the reagent per 15 millions of cells in 30 ml of the above medium. (dna-pkinhibitor.com)

In nature, plasmids often carry genes that benefit the survival of the organism and confer selective advantage such as antibiotic resistance. (wikipedia.org)
While chromosomes are large and contain all the essential genetic information for living under normal conditions, plasmids are usually very small and contain only additional genes that may be useful in certain situations or conditions. (wikipedia.org)
Smaller plasmids make use of the host replicative enzymes to make copies of themselves, while larger plasmids may carry genes specific for the replication of those plasmids. (wikipedia.org)
Many of the genes carried by a plasmid are beneficial for the host cells, for example: enabling the host cell to survive in an environment that would otherwise be lethal or restrictive for growth. (wikipedia.org)
We performed extensive functional characterization of diterpenoid phytoalexin biosynthetic genes in rice, and found that the genes for the biosynthesis of the major diterpenoid phytoalexins, phytocassanes and momilactones, are clustered on chromosomes 2 and 4, and that their expression is coordinately induced in rice cells after elicitation. (go.jp)
Transfection into mammalian cells is followed by screening for cell lines permanently expressing specific genes through their incorporation into the cellular genome. (welgen.com)
The technology utilizes fluorogenic oligonucleotide signaling probes and flow cytometry to detect and isolate individual living cells expressing one or more genes. (genelink.com)
Originating at The Rockefeller University, Chromovert Technology is a cell engineering tool for rapid production of stable cell lines expressing one or more genes. (genelink.com)
demonstrated endothelial-cell-specific delivery of siRNA by targeted Anti-VCAM-1 SAINT-O-Somes in vivo , resulting in downregulation of inflammatory genes in inflamed mouse kidney. (synvoluxproducts.com)

Once these high producers are identified, the cell lines and/or the proteins produced by the cells are validated. (labroots.com)
The strategic partnership with Selexis allows Avacta to access the technology and know-how to develop high-expressing CHO cell lines as well as the extensive experience of developing a range of Fc fusion proteins for clinical manufacturing. (kbibiopharma.com)
Few balancing acts are as delicate as the optimization of a cell expression system, especially if the expression system is to yield recombinant proteins destined for therapeutic use. (genengnews.com)
Further, there were no significant changes in metabolites or increases in biomass such as cell debris and host proteins. (genengnews.com)
We demonstrate that recombinant LHCGR proteins which include the hormone-binding domain are secreted from transfected cells and that natural LHCGR is also secreted from human placental explants. (biomedcentral.com)
LHCGR recombinant proteins representing varying lengths of the N-terminal extracellular domain were expressed in Chinese Hamster Ovary cells in suspension culture. (biomedcentral.com)
When CRISPR genome editing was introduced in 2009, plasmids were generously made available by the labs where they were developed so that other researchers around the world could use them to express Cas proteins in CRISPR experiments. (idtdna.com)
It had been shown that certain altered PrP proteins caused the death of brain cells by allowing excessive electrical charges to cross the membranes of the cell. (elifesciences.org)
In this case, production from stable cell lines is highly advantageous since it allows easy generation of larger quantities of recombinant proteins and antibodies in measures of up to several grams per batch. (invivo.de)
All these combined with a highly optimized process for the selection of best-producing cell pools and screening for high-producing monoclonals, lead to cost-optimized production of target proteins in large quantities. (invivo.de)

This technique has been used to obtain both transient 1 and stable 2 transfections in a wide variety of cell types. (sigmaaldrich.com)
Importantly, it has also been shown that plasmid transfections, specifically in CRISPR experiments, can cause cytotoxicity in embryonic stem cells [ 3 ], which could be of concern to many researchers using this cell type. (idtdna.com)
Even with cells that can tolerate plasmid transfections, there are other problems associated with the use of plasmids for CRISPR editing, such as complications in the timing of CRISPR experiments. (idtdna.com)
Compatible with adherent and suspension cells, the animal-origin free Trans IT-X2® achieves both exceptional transient and stable transfections. (bioscience.co.uk)

Combined Numerical and Experimental Investigation of Localized Electroporation-Based Cell Transfection and Sampling. (uni-bielefeld.de)

In addition, clonal cell lines can be isolated and characterized if desired. (neb.com)
The goal is to produce, in a reasonable time frame, clonal cell lines that meet desired criteria for an application of interest. (genelink.com)
We can produce multiple clonal cell lines with various expression levels (low / medium / high expression). (altogenlabs.com)

Cambridge, UK, and Geneva, Switzerland, 20 June 2019 - Avacta Group plc (AIM: AVCT), the developer of Affimer ® biotherapeutics and research reagents, and Selexis, a pioneering life sciences company and global leader in mammalian cell line generation technology, have partnered to develop the Chinese hamster ovary (CHO) cell line that will be used to manufacture Avacta's first Affimer clinical candidate for first-time-in-human clinical trials. (kbibiopharma.com)
In addition, utilizing a stable mammalian cell line can make products more acceptable for clinical use. (altogenlabs.com)

By skipping the lengthy screening procedures and continuing with production directly from the cell pool, it is possible to reduce the costs and timeline of your project without compromising on the quality of the product. (invivo.de)

The Lipofectamine 3000 transfection protocol was developed to be easy to use, while still ensuring optimum performance and reliability in a wide panel of cell lines. (thermofisher.com)

B ) When pre-formed RNP is introduced by nucleofection into cells, the enzyme rapidly starts cutting targeted genomic DNA. (idtdna.com)

Spectacular increase in transfection efficiency and dramatic decrease in cell toxicity. (thermofisher.com)
We achieved a 3-fold enhancement in the transfection efficiency for adherent Chinese Hamster Ovary (CHO-K1) cells with this modified protocol. (elsevierpure.com)
Also, we report for the first time the successful photo-transfection of recently trypsinised cells with significantly high transfection efficiency when transfected with modified plasmid. (elsevierpure.com)

To facilitate cell culture optimization, Essential Pharma has developed Cell-Ess, a serum replacement that is free of animal-origin components. (genengnews.com)
After years of optimization work to counteract these bottlenecks, InVivo successfully established a proprietary platform for the development of stable cell lines using CHO (Chinese hamster ovary) cells as expression system. (invivo.de)

The manual screening methods traditionally used for cell line development are time-consuming and labor-intensive, creating a great demand for high-throughput, automated solutions for such efforts. (labroots.com)
Thousands of individual clones can then be isolated and expanded using automated cell culture methods. (genelink.com)
METHODS: H. pylori cells were treated with the Lla-Met compound, and the effect of the compound on the cell morphology, cell membrane permeability, and oxidation of the bacteria cell was assessed. (bvsalud.org)

Altogen Labs can provide development of stable cell lines in 28 days. (altogenlabs.com)

Three short hairpin RNA (shRNA) sequences will be selected and cloned into a pQuietU6 plasmid. (welgen.com)
CHROMOVERT® PROCESS a) cDNAs are subcloned for expression of mRNAs comprising 3' untranslated plasmid-encoded Chromo-Tag™ sequences for detection using fluorogenic oligonucleotide signaling probes. (genelink.com)

The 3 shRNA plasmids will then be co-transfected into 293 cells with an cDNA expression plasmid. (welgen.com)

In such cases, it is beneficial to switch to a recombinant approach and produce the antibody of interest in stable CHO cell pools. (invivo.de)
2. The adoption of antibody light-and heavy-chain single-plasmid expression and GS screening system eliminates the risk of antibiotic residue in the past antibiotic screening system. (trinomab.com)
We could design a strategy that selectively kills tumor cells based on a K V 10.1-specific antibody. (biomedcentral.com)

Chinese hamster ovary (CHO) is one of the most widely used non-human mammalian cell lines. (thermofisher.com)
Mammalian cell lines, especially those produced using immortalized lines like human embryonic kidney 293 (HEK 293) and Chinese hamster ovary (CHO) cells, are widely used in biological research, including in drug discovery and for biologics production. (genelink.com)
Understanding the scope of RGEN-mediated off-target effects in human and other eukaryotic cells will be critically essential if these nucleases are to be used widely for research and therapeutic applications. (cdc.gov)

Consider ribonucleoprotein (RNP) instead of CRISPR plasmids! (idtdna.com)
CRISPR plasmids expressing Cas enzymes and guide RNAs have been used for several years, but these plasmids have several limitations, including cytotoxicity and unpredictability of expression. (idtdna.com)
Ribonucleoprotein (RNP)-mediated CRISPR genome editing is more effective and avoids the problems associated with using plasmids. (idtdna.com)
These plasmids have been cited in numerous publications and have helped to advance understanding and applications of CRISPR-Cas genome editing. (idtdna.com)
Meanwhile, however, many discoveries have been made about the best approaches to CRISPR genome editing-and it turns out that CRISPR plasmids are often not the most efficient approach to use, due to undesired effects such as cytotoxicity and unpredictability, both of which are discussed below. (idtdna.com)
Transfecting a plasmid creates a waiting time before the CRISPR genome editing can begin. (idtdna.com)
Comparison of events after introducing CRISPR plasmid vs. ribonucleoprotein (RNP) into cells. (idtdna.com)
A ) When cells are transfected with a CRISPR plasmid, the plasmid is transported into the nucleus where transcription takes place. (idtdna.com)
TransI T-X2® from Mirus Bio is a low toxicity, dynamic transfection system for the superior transfection of plasmid DNA, siRNA, miRNA and CRISPR/Cas9 components into mammalian and primary cells. (bioscience.co.uk)
Recent work has demonstrated that clustered, regularly interspaced, short palindromic repeats (CRISPR)/CRISPR-associated (Cas) systems1-3 can serve as the basis of a simple and highly efficient method for performing genome editing in bacteria, yeast and human cells, as well as in vivo in whole organisms such as fruit flies, zebrafish and mice4-13. (cdc.gov)

The field induces a complex reaction between the membrane and the plasmid that is accumulated at the cell interface by electrophoretic forces. (uni-bielefeld.de)

Culture cells in standard serum-containing or serum-free medium appropriate for the cell type. (sigmaaldrich.com)
For example, with animal cell cultures, the trend is away from serum and toward defined media, that is, media uncomplicated by the presence of animal-origin components. (genengnews.com)
Use of serum to culture cells dates back nearly 70 years and continues to this day, albeit with less devotion. (genengnews.com)
We developed Cell-Ess ® , a novel lipid supplement produced under GMP conditions that serves as a serum-replacement, Dr. Elhofy explains. (genengnews.com)
The genetic presence of Bh in fecal samples was detected by real-time fluorescence quantitative polymerase chain reaction, the CD4+ T cell counts in venous blood was measured using flowcytometry, and the HIV VL in serum was quantified using fluorescence-based instruments. (bvsalud.org)

Note that the intensity of the fluorescence may vary, depending on the cell line and labeling substrate used. (neb.com)
Twenty four to 48 hours after transfection, begin selecting mammalian cultures in 600-1,200 µg/ml G418 (geneticin) depending on the cell line. (neb.com)
It is recommended that a kill curve be established for each cell line to determine optimal selection conditions. (neb.com)
Our faster and more predictive screening techniques may significantly increase speed of stable cell line development while improving quality and uniformity. (welgen.com)
Stable cell line development study report. (welgen.com)
In general, cell line production begins with the introduction of one or more plasmids encoding cDNAs of interest into a cell culture. (genelink.com)
Standard services include transfection of plasmid DNA (15 - 20 µg), drug selection of clonal cells, colony pick up, generation of a stable line, expression and functional screening (at least 10 passages), and final validation of construct expression by qRT-PCR and/or Western blot analysis. (altogenlabs.com)
The choice of vector depends on the specific application and the host cell line being used. (altogenlabs.com)
Select for stable expression: To generate a stable cell line, it is necessary to select for cells that have stably integrated and are expressing the transgene. (altogenlabs.com)
Characterize the stable cell line: The final step is to fully characterize the stable cell line by analyzing its growth rate, stability, and expression levels of the transgene. (altogenlabs.com)
IgG-displaying cells showing high reactivity to FcεRIα antigen were screened by the cell sorter, resulting in the establishment of a CHO cell line producing with higher reactivity than the parent C9 IgG. (elsevierpure.com)
However, developing a stable cell line can be a time and resource intensive process with several bottlenecks that need to be overcome in order to ensure a good yield output. (invivo.de)
We chose a well-established, cGMP-banked CHO cell line and combined it with our optimized cell culture processes using a high-performance, chemically-defined, animal component-free medium. (invivo.de)
In many cases, sufficient material can be obtained through production from stable CHO cell pools instead of going all the way to produce a stable cell line. (invivo.de)
When developing a stable cell line, the resulting cell pool then serves as starting point for single cell cloning and various screening processes, in which the clone with the highest productivity is identified and selected as basis for the stable cell line. (invivo.de)
To circumvent this problem and speed up research and development, a large part of the pre-clinical studies can be carried out using InVivo's CHO cell pool concept based on a well-established cGMP-banked cell line, which can later be transferred into GMP facilities. (invivo.de)
Es tablishment of the CHO/IR cell line was described previ ously. (liverx-receptor.com)

Angiogenesis is the formation of new blood vessels from the existing vasculature, and it is significantly increased during tumor development with the aim to supply nutrients and oxygen that support the high rate proliferation of tumor cells. (researchsquare.com)
1994. Mesothelial cell proliferation: A nonspecific response to lung injury associated with fibrosis. (cdc.gov)

CRH mRNA was detected in BE(2)C cells by real time PCR. (nel.edu)
Furthermore, the endogenous CRH mRNA expression was also increased when the BE(2)C cell were transfected with ERalpha and SUMO1 in contrast to the transfection with ERalpha alone. (nel.edu)

Differences in the biological effects of crocidolite asbestos and two glass fibres on epithelial lung cells. (cdc.gov)

The membrane-type IgG-expressing CHO cells were then converted to IgG-secreting cells by transfection with a plasmid coding Cre recombinase. (elsevierpure.com)
Labeling of transfected cells with a fluorescent SNAP-Cell substrate should show strong inner cell membrane fluorescence. (neb.com)
The plasmid must be present during the electric pulse if it is to be transferred across the membrane into the cytoplasm. (uni-bielefeld.de)
Only the localized part of the cell membrane brought to the permeabilized state by the external field is competent. (uni-bielefeld.de)
This leads to an insertion of the plasmid, which can then cross the membrane. (uni-bielefeld.de)

SLC22A17 ) in inner medullary collecting duct (IMCD) cells, which are present in vivo in a hyperosmotic/-tonic environment that activates canonical Wnt/β-catenin signaling. (biomedcentral.com)
In vivo experiments revealed that Lla-Met induced oxidation, DNA fragmentation, and phosphatidylserine ectropionation in H. pylori cells. (bvsalud.org)
Furthermore, SAINT-Vivo has been developed as an efficient in vivo delivery agent for DNA vaccines that have been shown to elicit potent T and B cell responses. (synvoluxproducts.com)
SAINT-O-Somes, invented by Synvolux and patent-pending, allow the targeted delivery of biomolecules in vitro and in vivo to specific cell types or tissues. (synvoluxproducts.com)

The procedure stated below is designed for the transfection of CHO cells with 1 µg/µL pSV40-CAT plasmid (diluted in sterile molecular biology grade water). (sigmaaldrich.com)
Cell transfection using femtosecond lasers is gaining importance for its proven ability to achieve selective transfection in a sterile and relatively non-invasive manner. (elsevierpure.com)
The emerging etiologic agent V. fl uvialis has caused cell lines by using sterile culture fi lters of the V. fl uvialis sporadic cases and outbreaks of diarrhea in several strains that were isolated as a sole pathogen. (cdc.gov)

DNA plasmids should be high-quality, ethanol-precipitated, resuspended in molecular biology grade water to a final concentration of 1 µg/µL. (sigmaaldrich.com)
These observations demonstrate that, by enabling intramolecular imaging under ambient conditions in whole intact cells, RESI closes the gap between super-resolution microscopy and structural biology studies and thus delivers information key to understanding complex biological systems. (nature.com)
There has been a tremendous explosion in the popularity of confocal microscopy in recent years, due in part to the relative ease with which extremely high-quality images can be obtained from specimens prepared for conventional fluorescence microscopy, and the growing number of applications in cell biology that rely on imaging both fixed and living cells and tissues. (olympusconfocal.com)

First, we in order to generate the relevant constructs for stable expression in CHO cells. (invivo.de)

To determine the effects of osmolarity/tonicity changes, Wnt/β-catenin and TLR-4 activation on Lcn2-R and Lcn2 expression and cell viability in rat primary IMCD and mouse (m)IMCD3 cells. (biomedcentral.com)
Cell viability/death was determined with MTT and LDH release assays. (biomedcentral.com)
Exposure of cells with endogenous or stably overexpressing Lcn2-R to apo-Lcn2 or LPS decreased cell viability. (biomedcentral.com)

Cells expressing antigen-specific human monoclonal IgGs with high affinity on the cell surface after normal folding and posttranscriptional modification were screened using a cell sorter. (elsevierpure.com)
This mammalian cell display method was applied to in vitro affinity maturation of monoclonal C9 IgG specific to the human high-affinity IgE receptor (FcεRIα). (elsevierpure.com)
When you use monoclonal antibodies from hybridoma cell lines that do not promise the expected yields, then this does not have to be the end of your project. (invivo.de)
The platform follows the internationally accepted "Quality by Design" (QbD) concept and the rigorous DoE test design method for process development, which can quickly complete the process development from monoclonal cell lines to 200L pilot scale, and obtain efficient, stable, and scalable production process. (trinomab.com)

This is typically done by treating the cells with a selective agent, such as a drug or antibiotic, that kills cells that have not integrated the transgene or are not expressing it at a sufficient level. (altogenlabs.com)

However, such resolutions do not directly translate to experiments in cells, and Ångström resolution has not been demonstrated to date. (nature.com)
Experiments showed that adding a large amount of normal PrP to the cells could prevent these effects. (elifesciences.org)

Synthetic plasmids are available for procurement over the internet. (wikipedia.org)
In basic research, as depicted in this image, Cell-Ess can enhance reproducibility because it is 100% synthetic and does not vary in composition from lot to lot. (genengnews.com)

These data illustrated that whereas CD47/SIRPα inhibition could potentiate tumor cell phagocytosis, CD40-mediated activation of a type I interferon response provided a bridge between macrophage- and T-cell-mediated immunity that significantly enhanced durable tumor control and rejection. (aacrjournals.org)
Tumor immunogenicity results from the processing, and subsequent presentation of tumor antigens released from dead or dying tumor cells on MHC molecules expressed on antigen-presenting cells (APC). (aacrjournals.org)
Currently, heat-sterilized, GMP-grade SAINT lipids are being used in a clinical trial at the Radboud University Medical Center (Nijmegen, the Netherlands) for dendritic cell vaccinations to boost graft-versus-tumor immunity. (synvoluxproducts.com)
After sensitization with cytotoxic drugs, scFv62-TRAIL induced apoptosis only in K V 10.1-positive cancer cells, but not in non-tumor cells, nor in tumor cells lacking K V 10.1 expression. (biomedcentral.com)

Plasmids can also provide bacteria with the ability to fix nitrogen. (wikipedia.org)
A recent study has implicated Lcn2 secretion from collecting duct (CD) α-intercalated cells as a bacteriostatic defense mechanism against pathogenic bacteria in the urine [ 14 ]. (biomedcentral.com)
Although Maraffini and colleagues7 recently performed a systematic investigation of Cas9 RGEN specificity in bacteria, the specificities of RGENs in human cells have not been extensively defined and, to our knowledge, bona fide off-target mutations induced by Cas9 have not been identified in any eukaryotic cell or organism. (cdc.gov)

Additionally, the truncated Lhcgr, misrouted the full-length Lhcgr in ER, thereby decreasing the cell surface expression of the mature functional receptor. (biomedcentral.com)

Plasmids are considered replicons, units of DNA capable of replicating autonomously within a suitable host. (wikipedia.org)

Additional fee will be charged for Western blot or cell-based assay of your interest. (welgen.com)
Here, we use a human cell-based reporter assay to characterize off- target cleavage of Cas9-based RGENs. (cdc.gov)

Later in 1968, it was decided that the term plasmid should be adopted as the term for extrachromosomal genetic element, and to distinguish it from viruses, the definition was narrowed to genetic elements that exist exclusively or predominantly outside of the chromosome and can replicate autonomously. (wikipedia.org)
A few types of plasmids can also insert into the host chromosome, and these integrative plasmids are sometimes referred to as episomes in prokaryotes. (wikipedia.org)

These linear plasmids require specialized mechanisms to replicate their ends. (wikipedia.org)
We use our method in a proof-of-principle demonstration to map the molecular arrangement of the immunotherapy target CD20 in situ in untreated and drug-treated cells, which opens possibilities for assessing the molecular mechanisms of targeted immunotherapy. (nature.com)

1. HD-BIOP3 metrocyte platform based on CHO K1 suspension-cultured cell. (trinomab.com)

Calcium phosphate transfection is a commonly used method for the introduction of DNA into eukaryotic cells. (sigmaaldrich.com)

During embryonic morphogenesis, adhesion molecules are required for selective cell-cell interactions. (rupress.org)

For example, SAINT-O-Somes targeted to VCAM-1 and E-selectin have been demonstrated to selectively and functionally deliver siRNA into inflammation-activated endothelial cells in vitro . (synvoluxproducts.com)

Cells have been counted by trypan blue exclusion and fluorescence microscopy at 48 h post-transfection. (dna-pkinhibitor.com)

Indeed, research shows that transfecting some cell types with any plasmid may cause cell death [ 1 ], while some transfection reagents (e.g., lipids) used for plasmid transfection are themselves toxic to cells [ 2 ]. (idtdna.com)

Chinese hamster ovary cells overexpressing the insulin receptor were a present from Dr. Feng Liu. (liverx-receptor.com)
The effect of these parameters was systematically investigated using the transient expression of the chloramphenicol acetyltransferase and the beta-galactosidase activities in Chinese hamster ovary cells. (uni-bielefeld.de)

Anatomy9

Organisms18

Chemicals and Drugs44

Analytical, Diagnostic and Therapeutic Techniques and Equipment26

Phenomena and Processes73

Information Science4

Health Care1

Gene26

Protein27

Transient transfection4

HEK2931

Vectors2

Transfect8

EGFP1

HeLa1

Bacterium1

Reagent6

Genes9

Proteins10

Transfections4

Electroporation1

Clonal cell lines3

Mammalian cell line2

Timeline1

Lipofectamine1

Genomic1

Transfection efficiency3

Optimization2

METHODS3

Development of stable cell lines1

Sequences2

CDNA1

Antibody3

Widely3

CRISPR10

Induces1

Serum5

Line17

Proliferation2

MRNA2

Epithelial1

Membrane5

Vivo4

Sterile3

Biology3

Constructs1

Viability3

Monoclonal4

Antibiotic1

Experiments2

Synthetic2

Tumor4

Bacteria3

Functional1

Suitable host1

Assay2

Chromosome2

Mechanisms2

Suspension1

Eukaryotic cells1

Embryonic1

Endothelial1

Post-transfection1

Toxic1

Chinese2