靶向CDC42基因小干扰RNA分子的制备  

Preparation of siRNA Targeting for CDC42 Gene

作　　者：邵圣文[1] 薛利军[2] 周洪昌[1] 段劲[1] 王洁[2] 卢添宝[1] 徐菊玲[1] 

机构地区：[1]湖州师范学院医学院创新实验室,浙江313000 [2]解放军南京军区南京总医院血液肿瘤科

出　　处：《中国慢性病预防与控制》2011年第1期69-71,共3页Chinese Journal of Prevention and Control of Chronic Diseases

基　　金：国家自然科学基金(30800988)

摘　　要：目的制备靶向CDC42基因的小干扰RNA分子(siRNA)并检测其对CDC42基因表达的抑制效果。方法针对CDC42基因的不同区域设计4条siRNA分子,体外转录法合成siRNA分子,将siRNA与CDC42基因真核表达载体共转染HEK293T细胞,Westernblot法测定CDC42蛋白含量。结果合成了siR1、siR2、siR3、siR4共4条siRNA分子,siR3对CDC42基因表达的抑制率为95%,siR1、siR2以及siR4对CDC42基因表达的抑制作用不明显。siR3对HEK293T细胞生长无明显影响。结论体外转录法制备的siR3分子能够高效特异地抑制CDC42基因表达。Objective To prepare small interfering RNA （siRNA） targeting for CDC42 gene and determine the inhibition effect of siRNA on CDC42 gene expression. Methods Four chains of siRNA targeting for different coding region in CDC42 gene were designed and made by transcription in vitro, After siRNA and CDC42 eukaryotic expression vector were co-transferred into HEK 293T cells, CDC42 protein expression was detected by Western blot. Results Four chains of siRNA （siR1, siR2, siR3, siR4） targeting for CDC42 gene were obtained by transcription in vitro. The inhibition rate of siR3 on CDC42 gene expression was 95%, and siR1, siR2 or siR4 showed no obvious inhibition effect on CDC42 gene expression. The siR3 molecule didn＇t have obvious effect on the growth of HEK 293T cells. Conclusion The siR3 molecule prepared by transcription in vitro, is able to efficiently and specifically inhibit CDC42 gene expression.

关 键 词：CDC42基因 小干扰RNA 体外转录 

分 类 号：R372[医药卫生—病原生物学]