基础医学与临床2012，Vol.32Issue(4)：381-385,5.
293-siDrosha稳定细胞系模型的建立及microRNA相关功能的检测
Establishment of 293-siDrosha stable cell line and its potential use in the study of microRNA functions
摘要
Abstract
Objective To construct Drosha-1 deficient 293 stable cells (293-siDrosha) and to study microRNA processing by using the established 293 stable cells. Methods The pSicoR-human Droshal plasmid DNAs were transfected into 293 cells and selected for neomycin resistant clones with 1 200 g/mL G418 for 2 ~ 3 weeks. The mRNA and protein expressions of Drosha were detected by RT-PCR, real-time PCR and Western blotting. The expressions of has-miR-491-5p in 293 cells, uncloned 293-siDrosha cells (293-siDrosha mixed) and the cloned 293-siDrosha cells (293-siDrosha #3 cells) were detected with poly (A) real-time RT-PCR. Results Compared with the control cells, the mRNA and protein expression of human Droshal in cloned 293-siDrosha cells was dramatically decreased. The expression levels of has-miR-491-5p in the cloned 293-siDrosha cells have about 120 fold reduction as compared with that of the control cells. Conclusions 293-siDrosha stable cells were successfully constructed. The cloned 293-siDrosha cells may be used as a model system for the study of microRNA functions.关键词
siDrosha/microRNAs/293细胞Key words
siDrosha/ microRNAs/ 293 cells分类
医药卫生引用本文复制引用
郭蓉,刘力..293-siDrosha稳定细胞系模型的建立及microRNA相关功能的检测[J].基础医学与临床,2012,32(4):381-385,5.基金项目
国家自然科学基金(81171944) （81171944）
2010年中国医学科学院基础医学研究所所院长基金(2010PYZ17) （2010PYZ17）
