广西医科大学学报2018，Vol.35Issue(5)：603-607,5.DOI:10.16190/j.cnki.45-1211/r.2018.05.005
利用CRISPR/Cas9技术构建ATF4基因敲除的HepG2细胞株
Construction of ATF4 knockout HepG2 cell line by CRSPR/Cas9
摘要
Abstract
Objective:To construct ATF4 knockout HepG2 cells by using CRSPR/Cas 9 genome engineering technology.Methods:Two pairs of gRNAs targeting the ATF4 genes Exon 1 and 2 were designed for the functional domains of the ATF4 gene.The recombinant plasmid PX459-gRNA was constructed and transformed into competent cells DH5α.The recombinant plasmid was screened and sequenced.The effectiveness of the designed gRNA was confirmed by sequencing.PX459-ATF4-gRNA was transfected into HepG2 cells.After cloning and sequencing,The ATF4 knockout HepG2 cells were obtained and the expression of ATF4 was detected by western blotting.Results:The knockout plasmid was successfully constructed.Compared with negative control group,ATF4 gene was not expressed in the transfected PX459-ATF4-gRNA plasmid group.Conclusion:The ATF4 knockout HepG2 cell line was successfully constructed by using CRSPR/Cas 9 system,which lays the foundation for further studies of the effect and mechanisms of ATF4 in liver cancer.关键词
ATF4/CRSPR/Cas9系统/基因敲除/肝癌Key words
ATF4/CRISPR/Cas9 system/gene knockdown/heptocellular carcinoma分类
医药卫生引用本文复制引用
易桥勇,王秋雁,李天宇..利用CRISPR/Cas9技术构建ATF4基因敲除的HepG2细胞株[J].广西医科大学学报,2018,35(5):603-607,5.基金项目
国家自然科学基金资助项目(No.81760454) （No.81760454）
