用体外转录法制备小干扰RNA抑制Hela细胞CUEDC2基因表达  

Knockdown CUEDC2 in Hela Cells by Using in Vitro-transcripted siRNA

作　　者：李涛[1] 高彦飞[1] 巩伟丽[1] 靳宝锋[1] 

机构地区：[1]国家生物医学分析中心,北京100850

出　　处：《科学技术与工程》2010年第15期3574-3576,3581,共4页Science Technology and Engineering

基　　金：国家自然科学基金(30871275)资助

摘　　要：CUEDC2是一个广泛表达于各种真核生物、功能尚不明确的基因。利用一种改进的体外转录方法获得包含CUEDC2基因靶序列的长链RNA,进而通过RNA酶H切割引导序列形成成熟siRNA。转染Hela细胞并进行免疫印迹检测。结果显示,运用此方法得到的siRNA能够有效地抑制细胞中内源CUEDC2的表达,此项方法为CUEDC2的深入功能分析奠定了基础。A modified in vitro-transcripted method is used to produce siRNA for down-regulating the expression of CUEDC2,a widely expressed gene in all kinds of eukaryocyte.Long strand RNAs containing the leading sequence and the targeting sequence of CUEDC2 were obtained by an in vitro transcriptional system,the mature siRNAs were produced from the transcriptional RNA precursor by RNase H degradation.The siRNA were transfected into Hela cells and detected with western blot,the result shows that the expression of endogenous CUEDC2 is efficiently knocked down by siRNA,so The functions of CUEDC2 based on this study be could further analysed.

关 键 词：体外转录 小干扰RNA CUEDC2 

分 类 号：Q344.13[生物学—遗传学]