利用RNA干扰抑制HeLa细胞中CTCF基因的表达  

Inhibition of CTCF Gene Expression in HeLa Cells by RNA Interference

作　　者：何莉[1] 朱旭东[1] 党颖[1] 李涛[1] 黄培堂[1] 

机构地区：[1]军事医学科学院生物工程研究所

出　　处：《生物技术通讯》2006年第4期506-508,共3页Letters in Biotechnology

基　　金：国家科技攻关课题(2004AA002100)

摘　　要：目的:CTCF是一种多功能的转录因子,参与启动子调控、绝缘子功能和遗传印记等过程。利用RNA干扰技术抑制CTCF在体细胞中的表达。方法:以CTCF为靶基因,利用ShortCutRNaseⅢ切割体外转录的长双链RNA制备esiRNA,转染HeLa细胞,用RTReal-TimePCR检测CTCF的mRNA水平,用免疫印迹检测CTCF水平。结果:转染CTCFesiRNA后,细胞内的CTCFmRNA被抑制了70%左右,CTCF水平明显下降。结论:CTCF的esiRNA可以明显抑制CTCF在HeLa细胞中的表达,从而为进一步研究CTCF的未知功能打下了基础。Objective： CTCF is a multifunctional transcription factor, which plays many roles in promoter regulation, insulator function and genetic imprinting. To inhibit CTCF expression in mammalian cells by RNAi technology. Methods： The CTCF esiRNAs were prepared by cutting in vitro transcripted long dsRNA with ShortCut RNase Ⅲ, then were transfected into HeLa cells, and the CTCF mRNA and protein level were detected. Results： Both CTCF mRNA and CTCF protein level decreased dramatically after transfecting CTCF esiRNA in HeLa cells. Conclusion： CTCF expression in mammalian cells was successfully inhibited by RNAi technology. It was a primary experiment to further explore unknown functions of CTCF.

关 键 词：CTCF 转录因子 RNA干扰 基因表达 

分 类 号：Q75[生物学—分子生物学] Q78