【摘要】 背景与目的:双链RNA特异性地降解相应序列的mRNA,从而导致转录后水平的基因沉默。本研究旨在探讨绿色荧光蛋白(greenfluorescenceprotein,GFP)的短双链RNA(shortinterferingRNAs,siRNAs)是否能抑制GFP在哺乳动物细胞中的表达。方法:用荧光显微镜和荧光分光光度计观察合成的GFPsiRNAs对绿色荧光蛋白表达基因pEGFPC2在HeLa细胞中表达的影响,同时观察长链GFPdsRNA、Cx26siRNAs和长链Cx26dsRNA对荧光蛋白表达的影响,并通过流式细胞仪观察上述不同种类的dsRNA对细胞凋亡的影响。结果:GFPsiRNAs能特异性抑制GFP基因在HeLa细胞中的表达,而Cx26siRNAs对荧光蛋白的表达没有影响。长链双链RNA可以诱导细胞的凋亡,而siRNAs对细胞的凋亡无影响。结论:siRNAs能在哺乳动物特异性抑制基因的表达,该研究将为利用siRNA作为一种治疗手段提供一定的理论依据。更多还原

【Abstract】 Background and Objective:Double-stranded RNA(dsRNA)has been shown to interfere the function of specific genes in various species.T he current study was designed to investigate whether short interfering RNAs (siRNAs )of green fluorescence protein(GFP)can inhibit the expression of GFP in mammalian cells.Methods:HeLa cells were grown in standard conditions and transfecte d with pEGFPC2and long dsRNA or siRNAs.Fluoroscopy and fluoroscent spectrophotometer were used to evaluate impact of GFPds RNA,Cx26siRNAs ,and longer double-strand RNAs on GFP expression,and flow cytometry was u sed to evaluate the cell apoptosis.Results:Synthetic siRNAs can inhibite specific expression of GFP gene in human HeLa c ell lines and siRNAs have no such spec ific effect.Longer double-stranded but no siRNAs can induce cell apoptosis.Conclusion:These observations may open a path to ward the use of siRNAs as a new tool for researching gene function in mam malian cells and a gene-specific therapeutic tool.更多还原