摘要
目的构建联合靶向凋亡抑制基因Survivin和Livin的双重siRNA表达载体并鉴定其活性,为研究同时下调Survivin和Livin表达的协同效应奠定基础。方法选取已证实有效的分别靶向Survivin和Livin的siRNA序列,通过定向克隆技术构建同时编码2个siRNA分子的真核表达载体pSilencer2.1-Sur-Liv,将其转染HepG2细胞后检测Survivin和Livin mRNA水平的变化,同时转染表达DsRed2-SURVIVIN和GFP-LIVIN的Hela荧光报告细胞观察荧光强度变化。结果双重siRNA表达载体pSilencer2.1-Sur-Liv构建成功,转染HepG2后Survivin和Livin mRNA水平明显下降,转染Hela报告细胞后荧光强度变弱。结论联合靶向Survivin和Livin的双重siRNA表达载体构建成功,并能有效下调Survivin和Livin的表达水平。
Objective To construct a dual-siRNA expression vector targeting Survivin and Livin and identify its activity,and provide evidence for down-regulation of both genes simultaneously. Methods The published siRNA sequences targeting Survivin and Livin, respectively, were chosen to construct the dual-siRNA- expressed vector pSileucer2, l-Sur-Liv. This vector was then transteeted into HepG2 cells and the mRNA levels of these two genes were detected. This vector was also trausfected into Hela reporter cells, which expressed DsRed2-SURVIVIN and GFP-LIVIN,for examining the florescence. Results The recombinant vector pSilencer2, l-Sur-Liv was successfully constructed. After transfeeted with pSilencer2.1-Sur-Liv,the mRNA levels of Suruivin and Livin in HepG2 cells were obviously decreased and the florescence in Hela reporter cells was reduced. Conclusion The dual-siRNA-expressed vector targeting Survivin and Livin has been constructed successfully and may effectively suppress the expression of Survivin and Livin.
出处
《广东药学院学报》
CAS
2013年第3期314-317,共4页
Academic Journal of Guangdong College of Pharmacy
基金
广东省优秀博士论文作者资助项目(sybzzxm201051)
广东药学院科研启动基金(2007SMK02)
