As a leading provider of gene editing service, CRISPR/Cas9 PlatformCB is proud to offer gene knockout cell line generation service with high quality. Depending on the excellent platform and experienced scientists, our staff has succeeded in lots of gene knockout projects, ranging from tumor cell line to difficult-to-handle cell line. At CRISPR/Cas9 PlatformCB, we take the time to understand every case to help ensure you receive the edited cell line you desire. Our scientists will work with you closely to assist you in achieving your research goal.
Gene knockout cell line is a useful tool for research and manufacture. But it takes a long time to generate a knockout cell line. The most common method for gene editing is to use engineered nucleases, which can induce a double-strand break in genomic DNA. The induced double strand breaks are repaired through NHEJ or HR pathway, resulting in targeted mutations. To date, there are three widespread engineered nucleases for gene editing, ZFNs, TALENs and CRISPR/Cas9. As the latest technology, CRISPR/Cas9 are widely accepted for gene knockout and knockin. In CRISPR/Cas9 system, there are two key factors, Cas9 enzyme and guide RNA. Cas9 nuclease select the correct location on the genome by utilizing the sgRNA sequence to bond with base pairs on the host DNA. The sgRNA sequence, which is customizable and can be independently synthesized, is following a certain rule (PAM). Once it is transfected into cells, the Cas9 protein with the help of sgRNA finds the correct sequence and induced DNA breaks. The subsequent repair pathway is error prone and may induce indels.
Fig 1. An example of gene editing by CRISPR/Cas9
Procedure of generating knockout cell line
CRISPR/Cas9 PlatformCB offers one-stop knockout cell line generation service for our clients. Our custom cell line service begins with free consultation in which we will run a feasibility analysis for each case. Once the project begins, we will work closely with you and regularly update the project progress. CRISPR/Cas9 PlatformCB is your reliable partner for your research at all time.