Inhibition of intracellular hepatitis C virus replication by synthetic and vector-derived small interfering RNAs
Inhibition of intracellular hepatitis C virus replication by synthetic and vector-derived small interfering RNAs
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DOI：
10.1038/sj.embor.embor840
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发表时间：
2003-06-01
期刊：
影响因子：
7.7
通讯作者：
Mizusawa, H
中科院分区：
文献类型：
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作者：
Yokota, T;Sakamoto, N;Mizusawa, H
小干扰RNA（siRNA）通过RNA干扰有效地抑制基因表达。在这里，我们报告有效的抑制，合成和载体衍生的siRNA，丙型肝炎病毒（HCV）的复制，以及病毒蛋白质的合成，使用HCV复制子系统。siRNA被设计为靶向HCV基因组的5'非翻译区（5' UTR），其具有用于翻译整个病毒多蛋白的内部核糖体进入位点。此外，5'UTR是HCV基因组中最保守的区域，使其成为siRNA的理想靶标。重要的是，我们已经在5' UTR中确定了一个有效位点，在该位点上，用低至2.5 nM的siRNA浓度就可以实现对HCV复制的类似80%的抑制。此外，表达针对HCV的siRNA的基于DNA的载体也是有效的，这可能允许使用病毒载体将RNAi有效递送到体内肝细胞中。我们的研究结果支持使用基于siRNA的基因治疗来抑制HCV复制的可行性，这可能被证明在治疗丙型肝炎中是有价值的。
Small interfering RNAs (siRNAs) efficiently inhibit gene expression by RNA interference. Here, we report efficient inhibition, by both synthetic and vector-derived siRNAs, of hepatitis C virus (HCV) replication, as well as viral protein synthesis, using an HCV replicon system. The siRNAs were designed to target the 5' untranslated region ( 5' UTR) of the HCV genome, which has an internal ribosomal entry site for the translation of the entire viral polyprotein. Moreover, the 5' UTR is the most conserved region in the HCV genome, making it an ideal target for siRNAs. Importantly, we have identified an effective site in the 5' UTR at which similar to80% suppression of HCV replication was achieved with concentrations of siRNA as low as 2.5 nM. Furthermore, DNA-based vectors expressing siRNA against HCV were also effective, which might allow the efficient delivery of RNAi into hepatocytes in vivo using viral vectors. Our results support the feasibility of using siRNA-based gene therapy to inhibit HCV replication, which may prove to be valuable in the treatment of hepatitis C.