动物造模,CRISPR-Cas9 z-bio 2022-12-15 803

　　Purpose: Use the CRISPR/Cas9 system to knock out the MATP gene in mouse melanoma cell lines, and lay the foundation for the functional study of the MATP gene.

　　Method: Using , design specific primers for MATP, and link the primers to the pCAS9/gRNA1 vector. The positive vector was transfected into the mouse melanoma cell line B16F10, and the transfected monoclonal cell line was obtained by infinite dilution. The genomes of different cell lines were extracted, and the cell lines with MATP gene cut were further screened by sequencing, and the expression of MATP was verified by Western-blot method.

　　Result: Three cell lines with MATP gene knockout were successfully obtained. Western-blot results showed that the cell line did not express MATP protein.

　　Conclusion: Using pCAS9/gRNA1 vector, the MATP gene of B16F10 cell line can be knocked out.