Single Gene Knock Out Cell Line Generation

CRISPR/Cas9 is a revolutionary technology in the field of gene engineering. It has expedited the course of biopharmaceutical research and drug discovery, by providing scientists with a versatile tool to knockout any gene, in any cell and without introducing foreign DNA. The benefits of CRISPR/Cas9 over previous forms of gene editing, such as TALENs and zinc finger nuclease (ZFN), are that it is much simpler to implement and has higher efficiency at performing bi-allelic gene modifications.

Creative Biogene is experienced to provide a full CRISPR/Cas9-based single gene knockout service using any mammalian cell line and targeting any gene. Our scientists are experts at performing gene knockout with CRISPR/Cas9, from designing gRNA constructs to transfection and single clone generation of a wide range of cells, including difficult-to-transfect and tumor cell lines.

Our services include:

Host cell characterization

Optimize transfection methods of the host cell: Try all kinds of methods, including liposome-mediated transfection, electroporation, lentivirus-mediated transduction, et al.

Test the minimal absolute lethal dose of antibiotic concentration.

Test the clonability of the host cell: Determine if monoclonal cells are cultured.

gRNA design and KO vector construction

gRNAs are designed in common exons of different transcriptional products, generally.

Targets can destroy important domains of the protein and all the alternatively spliced transcripts.

Transfection

Transfect cells with the optimal transfection methods.

Single cell screening

Select cell by MACS, FACS or antibiotic selection and screen gene-edited cell clones.

Expand the engineered cells.

Validation

Evaluate the target stable cell clones via one or several assays such as Western Blot, ELISA, real-time PCR, or reporter assays etc.