Gene Knockout Stable Cell Lines

Product Details

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

• One-stop service: Customized cell generation project was managed by experienced scientists throughout the process, from construction design to drug screening.
• Experienced: Creative Biogene has professional research team with experience of many years in the field of molecular and cell biology.
• Genetic stability: Our knockout stable cell lines has been rigorously tested and validated.
• High Quality: Our knockout stable cell lines have no mycoplasma contamination.

Gene Knockout Stable Cell Line Product List

Application

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:

• Studying gene functions: One of the main applications of knockout cells is to decipher the function of genes of interest. By deleting the gene, researchers can evaluate its role in various biological processes, such as cell division, differentiation or organismal development. This knowledge is critical to understanding the underlying mechanisms of disease pathogenesis.
• Antibody specificity verification: To validate antibody specificity, gene knockout cells are utilized to confirm the absence of signal in immunostaining or western blotting experiments. The knockouts are generated by modifying the target gene within the cell's DNA using advanced techniques like CRISPR-Cas9 or homologous recombination. After the removal of the target gene, the resulting cell line lacks the corresponding protein on its surface or within its intracellular compartments. By comparing the staining or blotting results with control cells that possess the target gene intact, they can determine if the antibody recognizes its intended target accurately.
• Target validation and drug discovery: One of the key challenges in drug discovery is identifying appropriate drug targets. Gene knockout cells can facilitate target validation to assess the necessity of a specific gene as a therapeutic target. By deleting potential target genes, researchers can assess the impact on cell viability or disease progression. This information can help identify promising drug targets and develop effective treatment strategies.
• Establishment of disease models: Knockout cells can be used to model various genetic diseases. By disabling specific disease-related genes, researchers can study how the loss or dysfunction of these genes contributes to the development of the disease. This approach helps reveal the molecular mechanisms of many genetic diseases, including cancer, neurodegenerative diseases, and cardiovascular diseases.
• Develop treatment tools: In the past few years, researchers have conducted a series of gene-editing clinical trials, such as generating more effective CAR-T cells to treat cancer, and knocking out the erythroid-specific enhancer of BCL11A to upregulate gamma globulin in autologous erythroid HSCs as a potential treatment for sickle cell disease and beta-thalassemia. So far, there are already some cases of ongoing clinical trials using the method of knocking out the PD-1 gene of autologous T cells to treat cancer, including prostate cancer, esophageal cancer, and renal cell cancer. These trials are considered proof-of-concept studies for applying in vitro CRISPR/Cas9 gene knockout technology to cancer treatment.

Case Study

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)

FAQ