Stassen Liesel, Huismans Henk, Theron Jacques
Department of Microbiology and Plant Pathology, University of Pretoria, Pretoria 0002, South Africa.
Virus Genes. 2007 Dec;35(3):777-83. doi: 10.1007/s11262-007-0162-4. Epub 2007 Sep 13.
RNA interference (RNAi) is the process by which double-stranded RNA directs sequence-specific degradation of homologous mRNA. Short interfering RNAs (siRNAs) are the mediators of RNAi and represent powerful tools to silence gene expression in mammalian cells including genes of viral origin. In this study, we applied siRNAs targeting the VP7 gene of African horse sickness virus (AHSV) that encodes a structural protein required for stable capsid assembly. Using a VP7 expression reporter plasmid and an in vitro model of infection, we show that synthetic siRNA molecules corresponding to the AHSV VP7 gene silenced effectively VP7 protein and mRNA expression, and decreased production of infectious virus particles as evidenced by a reduction in the progeny virion titres when compared to control cells. This work establishes RNAi as a genetic tool for the study of AHSV and offers new possibilities for the analysis of viral genes important for AHSV physiology.
RNA干扰(RNAi)是双链RNA指导同源mRNA进行序列特异性降解的过程。小干扰RNA(siRNA)是RNAi的介导因子,是沉默哺乳动物细胞中基因表达(包括病毒起源基因)的有力工具。在本研究中,我们应用了靶向非洲马瘟病毒(AHSV)VP7基因的siRNA,该基因编码稳定衣壳组装所需的结构蛋白。使用VP7表达报告质粒和体外感染模型,我们发现与AHSV VP7基因对应的合成siRNA分子有效沉默了VP7蛋白和mRNA表达,并减少了感染性病毒颗粒的产生,与对照细胞相比,子代病毒粒子滴度降低证明了这一点。这项工作确立了RNAi作为研究AHSV的遗传工具,并为分析对AHSV生理学重要的病毒基因提供了新的可能性。