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Cas9 gene edited cell line construction

Basic Information

　　The CRISPR/Cas9 system consists of a single guide RNA (sgRNA) and a Cas9 protein with endonuclease activity. The technical principle involves two basic processes: sgRNA-guided Cas9 targeting DNA cleavage and DNA repair. The advantages of CRISPR/Cas9 technology are the ability to permanently alter the genomic DNA sequence and the ability to edit non-coding DNA regions. It also allows editing at multiple locations simultaneously.
　　Cell lines constructed by knocking out the target gene using Cas9 genome editing technology can be used in gene function research, disease model construction, gene therapy, and other fields, which has broad application prospects.

Services We Provide

Construction of Cas9 genome editing cell lines

Procedure of Services

　　（1） sgRNA Design and Vector Construction
　　This procedure includes target gene analysis, target design, and vector construction.
　　（2）Transfecting 293T cells with sgRNA screening, Cas9/gRNA and homologous recombination single-stranded donor.
　　（3） Monoclonal cell line screening
　　 Observe the fluorescence rate of cell transfection. Once the fluorescence rate meets the criterion, perform a limiting dilution of the cells into a 96-well plate at a single-cell level. After 24 to 48 hours, the single clones (GFP or RED fluorescent cells) were labelled and expanded for further testing.
　　（4） qPCR analysis and sequencing verification.
　　 Extract the genomic DNA from transfected cells and amplify the genomic sequences near the target site through PCR. Positive knockout clones were identified through sequencing.
　　（5） Cell expansion and seed preservation of the target cells.
　　（6）Delivery of experimental results
　　Monoclonal cell line of target gene knockout cells: 1*106 or 2*106 of cells.

Strengths and Features

　　（1）One stop integrated services
　　We provide one-stop service for CRISPR/Cas9 gene knockout and point mutation cell line construction, which effectively help our customers to control the costs of development.
　　（2）Quality assurance
　　We ensure that our cell strains are free from mycoplasma and chlamydia infections through strict quality control measures.
　　（3）Professional technical guidance
　　Providing professional cell culture and Cas9 technology guidance.
　　（4） After-sales service
　　We provide a 2-week no-obligation after-sales service upon receipt of the cells. Knockout cells are preserved for long-term use by the customers.

Representative Case

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