中国比较医学杂志2017，Vol.27Issue(4)：52-55,4.DOI:10.3969.j.issn.1671-7856.2017.04.009
利用CRISPR-Cas9系统敲除小鼠黑色素瘤细胞系MATP基因的初步研究
A preliminary study on the MATP gene knockout in a mouse melanoma cell line using CRISPR-Cas9 system
摘要
Abstract
Objective To knockout the MATP gene of mouse melanoma cell line B16F10 using CRISPR/Cas9 system,and to lay foundation for the functional study of MATP gene.Methods Specific primers of MATP were designed according to the report in http://crispr.mit.edu/ website.The primers were linked to pCAS9/gRNA1 vector.Then the positive vector was transfected into mouse melanoma B16F10 cells,and monoclonal cell lines were obtained by the infinite dilution method.After the genomes of different monoclonal cell lines were extracted and sequenced,the cell lines with MATP gene cleavage were screened,and the expression of MATP in these cell lines was verified by Western-blot analysis.Results Three MATP gene knockout cell lines were successfully obtained.The western-blot results showed that the cell lines did not express MATP protein.Conclusions The knockout of MATP gene in B16F10 cell line can be successfully achieved using the pCAS9/gRNA1 vector.关键词
CRISPR-Cas9/黑色素瘤细胞/MATP/研究Key words
CRISPR-Cas9/Melanoma cells/MATP/Mouse melanoma cell lines分类
医药卫生引用本文复制引用
殷慧慧,孙兆增,李丹,李钰,孙菲,董施施,孔江峰,王洪宝,曾林,法云智..利用CRISPR-Cas9系统敲除小鼠黑色素瘤细胞系MATP基因的初步研究[J].中国比较医学杂志,2017,27(4):52-55,4.基金项目
国家自然科学基金31272387,全军实验动物专项课题SYDW〔2014〕006. （）
