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Original article link: HeYuan Bio – Knockout Cell Line Construction
Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 is an adaptive immune defense mechanism that has evolved over long periods in bacteria and archaea. The CRISPR/Cas9 system can be used for genome editing across a wide range of organisms. It recognizes target sequences via a small guide RNA (sgRNA) and employs the Cas9 nuclease to cleave DNA at a predetermined site within the recognition sequence, generating a double-strand break. Subsequent error-prone repair by the cell leads to random mutations. With its high mutation efficiency, flexibility, simplicity, short experimental timelines, and low cost, the CRISPR/Cas9 system has been widely adopted in genetic‑editing research. Its applications include the generation of gene‑knockout cell lines and the study of gene‑knockout animal models.
The company offers the following project services: CRISPR/Cas9 experimental protocol design, construction of CRISPR/Cas9 gene knockout vectors, viral packaging of CRISPR/Cas9 gene knockout vectors, and generation of CRISPR/Cas9‑mediated gene knockout cell lines.
Service Features:
1. To generate a custom gene-knockout cell line, simply provide the gene name;
2. Rapid generation of effective sgRNAs and high gene-editing efficiency;
3. Short experimental duration;
4 CRISPR/Cas9 lentiviral packaging services are available upon request.
Operating Procedure
1 Design of sgRNAs targeting the desired sequence, vector construction, and activity validation
2 Co-transfection of the target cell line, followed by preliminary screening of mixed clones.
3. Plating of single clones, screening of single clones, and genotyping of cells
4 Expansion of the Positive Cell Line
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