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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...

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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.

Advantages:

  • One-stop service is provided from design of gRNAs, cell transfection, preparing of single clones to sequencing analysis, only needing customers to provide target genes information and the cell line of interest.
  • Our CRISPR gene knockout technology applies to targets of any genes and mammalian cells.
  • Professional researchers have rich cell culture experience and plentiful CRISPR genome editing experience.
  • The detailed project reports will be delivered to customers after services are finishes.
  • Cell lines can be made from nearly any cell type, including cells that are difficult to transfect and grow incl. primary cells.

Single Gene Knock Out Cell Line Generation Service Case

Knock out the Fut8 gene in CHO-K1 cells

1. Genomic sequencing results of Fut8 gene KO cell lines

2. qPCR analysis of Fut8 gene KO cell lines

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