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The CRISPR interference (CRISPRi) system is a modified version of the CRISPR/Cas9 gene-based technology that uses synthetic programmable transcription factors to initiate endogenous inhibition of genes. In mammalian cells, the expressing of dCas9 (a nuclease-deficient Cas9 only retaining DNA binding activity) fused with a transcriptional repressor, and a 20 nt guide sequence of gRNA molecules complementary to the target promoter region in the genome, will lead to transcriptional repression of the corresponding gene.
CRISPR/Cas9 PlatformCB has successfully implied CRISPR-mediated gene transcriptional inhibition in a variety of cells, which facilitates further research on gene transcriptional regulation mechanisms, gene functions, and screening of drug targets. Based on our platform, we are able to offer you custom CRISPRi stable cell line services, from strategic design to stable cells. We guarantee three homozygous (or heterozygous) clones in which the modified line has the correct genetic mutation and three vials of each clone.
Strategy of CRISPRi is described as follows
The fusion protein of dCas9 and transcriptional repressor fusion (e.g. Krüppel-associated box, KRAB) domain is recruited to the transcription start site (TSS) of the endogenous gene specified by the gRNA to inhibit transcription.
Workflow of CRISPRi stable cell lines
➢ Design an CRISRP interference strategy based on your needs
➢ Design and synthesize specific gRNA
➢ Transfection of specific cell lines
➢ Screening monoclonal strain
➢ Identify monoclonal strains by PCR and sequencing, and screen the successfully edited cell lines
➢ Amplification, cryopreservation, delivery
We provide custom cell lines according to your requirements, our scientists have extensive experience with >500 distinct cell lines and are very familiar with their culture conditions and genetic backgrounds.
Alternative cell lines
As a professional gene editing company, CRISPR/Cas9 PlatformCB is specializing in building efficient systems and procedures to meet the needs and timelines of clients working in the CRISPR/Cas9 gene editing. We have successfully obtained thousands of CRISPR/Cas9 cell lines in >200 different cell lines, including easy-to-transfer cell lines and difficult-to-transfect cell lines. We guarantee our clients the most reliable and efficient research services to best match your research goals, and faster turnaround time, lower price. If you have any question, don’t hesitate to contact us.