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The CRISPR/Cas9 system is an adaptive immunity system in bacteria against bacteriophage invading. It has been developed as the most popular method for genome editing in recent years. Creative Biogene is offering a series of gene knockout cell lines developed by the CRISPR/Cas9 system. Our scientists have years of experience in performing gene editing with CRISPR/Cas9, from designing gRNA to transfection and single clone generation. These cell lines provide you with a convenient means to study gene functions.
Advantages
Gene knockout cells are an invaluable tool in molecular biology and genetics research. They have revolutionized our understanding of gene functions by allowing scientists to investigate the effects of specific gene deletions or mutations. These cells serve as valuable models to study gene function and reveal the impact of a particular gene on cellular processes and organismal development.
Applications for gene knockout stable cell lines include:
Case Study 1
Excessive fibrosis underlies the dysfunction of many critical organs. Activated myofibroblasts accumulate within pathological lesions in various fibrotic diseases, including patchy and interstitial fibrosis in progressive heart failure and cardiac hypertrophy. Therefore, attenuation of myofibroblast differentiation is expected to alleviate fibrosis. The researchers sought to find potential targets to mitigate fibroblast differentiation by analyzing transcription factors in human fibroblasts/myofibroblasts, as fibroblasts undergo transcriptomic changes during differentiation. Here, researchers report that a novel molecular target, transcription factor AP-2α (TFAP2A), reduces fibroblast transdifferentiation.
Figure 1. Deletion of TFAP2A gene significantly reduces TGF‐β1‐induced fibroblast differentiation. (The TFAP2A knockout cell line with NIH/3T3 fibroblasts (TFAP2A‐KO) was established using CRISPR/CAS9 technology through Creative Biogene, Shirley, NY.) (Ross G R, et al. 2019,)
Case Study 2
Congenital anomalies of the kidney and urinary tract (CAKUT) are a diverse group of developmental defects that occur in approximately 1:500 live births. In addition to its high morbidity, CAKUT is the most common cause of renal failure in children. The study aimed to perform a miRNA-centric analysis to identify and functionally interpret miRNAs most frequently affected by rare CNVs in CAKUT complemented by a CNV-centric analysis to estimate the overall burden of rare CNVs on miRNA genes in CAKUT. Based on the functional interpretation, another aim of this study was to experimentally confirm the impact of rare CNVs in CAKUT on candidate miRNA expression and subsequent changes in the mRNA levels of selected target genes.
Figure 2. The difference in the relative expression of target genes mRNA between MIR484+/− Knockout and HEK293 WT. (The human embryonic kidney cell line (HEK293 WT) and heterozygous MIR484 (GeneID: 619553) knockout HEK293 cell line (MIR484+/- KO) were obtained from Creative Biogene (Shirley, NY, USA).) (Mitrovic K, et al. 2022)
Q: What is a knockout cell?
A: A knockout cell refers to a type of genetically modified cell used in research to understand the function of specific genes or proteins. In a knockout cell, a specific gene or protein is intentionally deactivated, or "knocked out," through techniques like gene editing or gene disruption.
Q: What is the difference between knockdown and knockout cells?
A: Gene knockout refers to the complete elimination of a gene from an organism. Gene knockdown refers to the reduction of gene expression in an organism. It can only be achieved through genetic engineering technology. It can occur through chemical agents or genetic modification.
Q: How are your knockout cell lines generated?
A: Our knockout cell lines were generated using CRISPR/Cas9 technology. knockout cell lines were monoclonal and verified using Sanger sequencing.
Q: Do you test your knockout cell lines for mycoplasma?
A: Yes, the cell line bank is tested for mycoplasma. In addition, we conduct regular spot checks of our inventory.
Q: What is the biosafety level (BSL) of your cells?
A: Our knockout cell lines are divided into BSL1 and BSL2 based on cell line. Cell lines such as A549, HCT116, HEPG2, and MCF7 belong to BSL1. HEK-293T and HeLa cell lines are BSL2.
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