应用RNA干扰抑制目的基因在小鼠体内表达  

Suppression of gene expression in cells by RNA interference

作　　者：谢书阳[1] 王萍玉[1] 王桂兰[1] 白咸勇[1] 李有杰[1] 张公文 

机构地区：[1]滨州医学院生物化学与分子生物学教研室,山东烟台264003

出　　处：《中国现代医学杂志》2008年第1期17-20,共4页China Journal of Modern Medicine

基　　金：山东省教育厅基金(106L01);滨州医学院科研基金(BY2005KYQD19)

摘　　要：目的研究RNA干扰(RNAi)对体外和体内基因表达的沉默作用。方法以PCR方法从基因组DNA中克隆出依赖于RNA聚合酶Ⅲ的H1启动子,并用于驱动RNAi片段的合成;构建特异性针对目的基因(绿色荧光蛋白,GFP)的RNAi载体(Pi)。将RNAi干扰载体和GFP载体转染NIH3T3细胞,应用荧光显微镜观察、RT-PCR、流式细胞技术(FACS)分析上述细胞中RNAi对目的基因GFP表达的抑制效果,进一步在个体水平将RNAi载体注入小鼠体内,研究RNAi对GFP表达的抑制情况。结果RNAi能有效地使NIH3T3细胞中GFP表达量降低约60%以上,并能有效地抑制GFP在转基因小鼠体内的表达。结论RNAi技术能抑制目的基因在体内外的表达,是一种有效的基因治疗工具。[Objective] To evaluate whether RNA interference （RNAi） can silence gene expression in vitro and in vivo. [Methods] H1 promoter was amplified from human genome DNA for driving RNAi synthesis by PCR. RNAi vectors （Pi） for silencing the GFP expression were constructed. The GFP vector and RNAi vector （Pi） were then cotransfected into the NIH3T3, in which the silencing effect on GFP expression was investigated by fluorescence microscopic observation, RT-PCR and FACS analysis. Then RNAi plasmid was injected into GFP transgenic mice by tail vein to investigate the silencing effect of RNAi to GFP expression in vivo. [Results] RNAi could effectively reduce more than 60 percent of GFP expression in NIH3T3 cells as well as inhibit the GFP expression in GFP transgenic mice. [Conclusions] RNAi technology, which can inhibit gene expression in cells, is an effective tool for gene therapy.

关 键 词：RNA干扰 基因表达 基因沉默 基因治疗 

分 类 号：Q785[生物学—分子生物学] R-332[医药卫生]