Intracellular-diced dsRNA has enhanced efficacy for silencing HCV RNA and overcomes variation in the viral genotype
Intracellular-diced dsRNA has enhanced efficacy for silencing HCV RNA and overcomes variation in the viral genotype
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DOI：
10.1038/sj.gt.3302734
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发表时间：
2006-06-01
期刊：
影响因子：
5.1
通讯作者：
Kohara, M.
中科院分区：
文献类型：
--
作者：
Watanabe, T.;Sudoh, M.;Kohara, M.
RNA干扰（RNAi）可用于抑制病毒在哺乳动物细胞中的复制，因此可能是一种强大的新的抗病毒治疗方法。小干扰RNA（siRNA）可能对RNAi有效，但在每种情况下都存在一些必须解决的技术问题，例如，预测有效的siRNA靶位点和靶向病毒群体中的异质序列。我们在这里表明，从长双链RNA（dsRNA）产生的切割siRNA是非常有效的诱导RNAi在HuH-7细胞携带丙型肝炎病毒（HCV）复制子，并可以克服HCV基因型的变化。然而，在哺乳动物细胞中，长dsRNA诱导干扰素应答并导致细胞死亡。在这里，我们描述了这种方法的改进，U6启动子驱动的表达长发夹RNA与多个点突变的正义链。这可以有效地沉默HCV RNA复制和HCV蛋白表达，而不会触发通常由dsRNA引起的干扰素应答或细胞死亡。总之，胞内切割的dsRNA有效地诱导RNAi，并且尽管HCV中突变率高，但它应该是沉默HCV RNA的可行的治疗策略。
RNA interference (RNAi) can be used to inhibit viral replication in mammalian cells and therefore could be a powerful new antiviral therapy. Small interfering RNA ( siRNA) may be effective for RNAi, but there are some technical problems that must be solved in each case, for example, predicting the effective siRNA target site and targeting heterogeneous sequences in a virus population. We show here that diced siRNA generated from long double-stranded RNA ( dsRNA) is highly effective for inducing RNAi in HuH-7 cells harboring hepatitis C virus (HCV) replicons and can overcome variations in the HCV genotype. However, in mammalian cells, long dsRNA induced an interferon response and caused cell death. Here we describe an improvement of this method, U6 promoter-driven expression of long hairpin-RNA with multiple point mutations in the sense strand. This can efficiently silence HCV RNA replication and HCV protein expression without triggering the interferon response or cell death normally caused by dsRNA. In conclusion, intracellular-diced dsRNA efficiently induces RNAi, and, despite the high rate of mutation in HCV, it should be a feasible therapeutic strategy for silencing HCV RNA.