携带甲胎蛋白基因小干扰RNA慢病毒载体的构建及其对靶
基因的沉默效率
程木华;李建芳;张峰;曾凤伟;谢良骏
【期刊名称】《中华生物医学工程杂志》 【年(卷),期】2012(018)005
【摘要】Objective To construct lentiviral vectors carrying small interference RNA (siRNA) of alpha-fetoprotein(AFP) followed by transfection into hepatocellular carcinoma cells and to assess the efficiency of AFP gene silencing.Methods The AFP-specific positive siRNA and non-specific negative siRNA were designed and constructed prior to insertion to the recombinant lentiviral vectors carrying green fluorescent protein (GFP) gene.This was followed by transfection into human hepatocellular carcinoma cells HepG2 and subsequent screening of cell strains with GFP expression,and were assigned into lentivirustransfected positive siRNA group and lentivirus-transfected negative siRNA group respectively.Furthermore,liposome-transfected positive siRNA group,liposome-transfected negative siRNA group,siRNA group which treated with AFP siRNA but without transfection reagent,and blank control group were established simultaneously.The efficiency
of
transfection
was
assessed
under
fluorescent
microscope,and the relative expression of AFP mRNA and protein in HepG2 cells was determined via fluorescent quantitative polymerase
携带甲胎蛋白基因小干扰RNA慢病毒载体的构建及其对靶基因的沉默效率
