中文摘要：

目的利用RNAi技术对HIV-1 vif基因的转录后水平进行下调，达到抑制vif蛋白表达的目的，为新的抗HIV治疗和预防研究提供理论基础。方法通过体外转录法合成siRNA，与转接有vif基因的表达质粒共同转染HEK293T细胞，荧光显微镜下观察干扰效果，并利用Real—time PCR和Western blot对HIV-1 vif分别从转录和表达水平进行验证。结果将pEGFP—N1-vif重组载体转染HEK293T细胞后，结果显示vif蛋白可以在细胞中高水平表达；针对vif设计的3段特异的siRNA可以有效且特异地降低vif蛋白的表达。结论RNAi技术可以下调HIV-1蛋白的表达，此技术可以作为一项新的治疗和预防HIV-1的方法用于进一步的研究。

英文摘要：

Objective In order to explore the potential effects of gene therapy for HIV-1 treatment and precaution, we studied the suppression of RNA interference on HIV-1 vif protein by directing vif to post-transcriptional level. Methods siRNAs, synthesized by transcription in vitro, and the plasmid expressing vif gene and EGFP were cotransfeeted into HEK 293T cell, to express the vif protein and investigate the suppression effect of siRNA on HIV-1 vif under fluorescent microscope. This effect was confirmed by real time PCR at transcription level and Western blot at expression level. Results HIV-1 vif protein exhibited high expression in HEK 293T cells after transfecting pEGFP- N1-HIV-1/vif plamid into HEK 293T cell alone; the three siRNAs designed in this study targeted to vif suppressed HIV-1 vif protein expression specifically. Conclusions The use of RNAi, a new gene therapy tool, was proved to be useful in inhibiting HIV-1 vif expression. This technique deserves further study as a potential therapeutic tool for HIV-1.