基于Smad7基因为靶序列的RNA干涉作用研究  

Study of RNA interference based on the target sequening of Smad 7 gene

作　　者：王菊萍[1] 王莹[2] 霍艳英[3] 郭蔼光[1] 徐勤枝[3] 张开泰[3] 

机构地区：[1]西北农林科技大学生命科学学院,陕西杨凌712100 [2]重庆医科大学基础医学院,重庆400016 [3]军事医学科学院放射医学研究所分子毒理研究室,北京100850

出　　处：《西北农林科技大学学报（自然科学版）》2004年第12期13-17,共5页Journal of Northwest A&F University(Natural Science Edition)

基　　金：国家重点基础研究发展规划973项目(G1998051207)

摘　　要：　为了探讨小干涉RNA(siRNA)对靶基因Smad7mRNA表达的阻断作用,采用体外转录方法制备Smad7基因的小干涉RNAs(siRNAs),用脂质体介导瞬时转染BERP35T 2肺癌细胞系,并采用Northernblot杂交检测靶基因mRNA的表达丰度。结果表明,根据Smad7编码区序列在体外成功地制备了针对2个不同靶序列的siRNAs;Northernblot杂交显示,在转染siRNA的BERP35T 2细胞中,不管是内源性的还是外源性的,Smad7mRNA的表达丰度均明显下降。说明Smad7基因编码区中542563bp及701722bp2个区域均是siRNA作用的有效靶序列,本研究设计并制备的siRNAs能有效抑制Smad7基因的表达。This research intended to investigate the suppression of siRNA to the expression of the mRNA of target gene Smad 7.The siRNAs of Smad 7 gene were prepared by in vitro transcription.BERP35T-2 lung cancer cell line was transient transfected with siRNAs and Smad 7 expression plasmid using Lipofectamine 2000 reagent.The expression abundance of target gene mRNA was tested with Northern blot hybridization.According to the coding sequence of Smad 7 gene,we successfully synthesized the siRNAs for two different target sequences in vitro.The result of Northern blot hybridization showed that the expression abundance of both endogenous and exogenous Smad 7 mRNA decreased evidently in BERP35T-2 cells transfected with siRNAs.Both regions of 542-563 bp and 701-722 bp in the coding region of Smad 7 gene are effective target sequences which can be acted by siRNA.The siRNAs designed and synthesized in our research can suppress the expression of Smad 7 gene effectively.

关 键 词：小干涉RNA Smad 7基因 体外转录 瞬时转染 靶序列 

分 类 号：Q786[生物学—分子生物学] R734.2[医药卫生—肿瘤]