Knockout Cell Line Generation    

As a leading provider of gene editing service, CRISPR/Cas9 Platform^CB 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 Platform^CB, 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, 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

1. Cell Line Evaluation:
   We will evaluate the cell line by experiments in several parts, e.g. clonality.
2. SgRNA design and synthesis (at least 3 sgRNA)
3. Select the most efficient sgRNA
4. Transfect the cells with sgRNA and Cas9
5. Single cell cloning and expansion
6. Screen for positive cell clones by sequencing
7. Prepare cells for delivery

Advantage

• Years of experience in CRISPR/Cas9 system
• Leading-edge equipment and platform
• Affordable price with high-quality service
• Fast turnaround time
• Multiplexed gene knockout cell is available

CRISPR/Cas9 Platform^CB 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 Platform^CB is your reliable partner for your research at all time.

Case Study

KO Cell Line Generation

For more details, please feel free to contact us.

Related Products

• Pre-made Knockout Cell Line
• Gene Knockout Cell lysate
• SuperKO sgRNA Vector