动物造模,黑色素瘤细胞 z-bio 2022-06-07 287

　　OBJECTIVE: To use the CRISPR/Cas9 system to knock out the MATP gene in mouse melanoma cell lines, laying the foundation for the functional study of the MATP gene.

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

　　RESULTS: Three MATP gene knockout cell lines were successfully obtained. Western-blot results showed that these cell lines did not express MATP protein.

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