Small interfering RNA targeting m2 gene induces effective and long term inhibition of influenza A virus replication.
Small interfering RNA targeting m2 gene induces effective and long term inhibition of influenza A virus replication.
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DOI：
10.1371/journal.pone.0005671
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发表时间：
2009-05-22
期刊：
影响因子：
3.7
通讯作者：
Zheng BJ
中科院分区：
文献类型：
--
作者：
Sui HY;Zhao GY;Huang JD;Jin DY;Yuen KY;Zheng BJ
RNA干扰（RNAi）为特异性抑制病毒感染提供了一种强有力的新手段。然而，sirna耐药病毒的选择是使用RNAi作为抗病毒治疗方法的一个主要问题。在这项研究中，我们利用带有h1短发夹RNA （shRNA）表达盒的慢病毒载体，将小干扰RNA （sirna）传递到哺乳动物细胞中。利用该表达增强型绿色荧光蛋白（EGFP）的载体作为替代标记，成功建立了稳定的shrna表达细胞系，并评估了合理设计的sirna对甲型流感病毒基因组保守区域的抑制效果。结果表明，一种靶向流感M2基因（siM2）的siRNA能有效抑制病毒复制。siM2不仅对H1N1病毒有效，而且对H5N1高致病性禽流感病毒也有效。除了抑制M2外，siM2还抑制NP mRNA的积累和蛋白表达。siM2的长期抑制作用已被证实，在流感准种中未观察到sirna抗性突变体的出现。综上所述，我们的研究表明M2基因可能是抗病毒治疗的最佳RNAi靶点。这些发现为开发基于rnai的人类流感病毒感染预防和治疗提供了有用的信息。
RNA interference (RNAi) provides a powerful new means to inhibit viral infection specifically. However, the selection of siRNA-resistant viruses is a major concern in the use of RNAi as antiviral therapeutics. In this study, we conducted a lentiviral vector with a H1-short hairpin RNA (shRNA) expression cassette to deliver small interfering RNAs (siRNAs) into mammalian cells. Using this vector that also expresses enhanced green fluorescence protein (EGFP) as surrogate marker, stable shRNA-expressing cell lines were successfully established and the inhibition efficiencies of rationally designed siRNAs targeting to conserved regions of influenza A virus genome were assessed. The results showed that a siRNA targeting influenza M2 gene (siM2) potently inhibited viral replication. The siM2 was not only effective for H1N1 virus but also for highly pathogenic avian influenza virus H5N1. In addition to its M2 inhibition, the siM2 also inhibited NP mRNA accumulation and protein expression. A long term inhibition effect of the siM2 was demonstrated and the emergence of siRNA-resistant mutants in influenza quasispecies was not observed. Taken together, our study suggested that M2 gene might be an optimal RNAi target for antiviral therapy. These findings provide useful information for the development of RNAi-based prophylaxis and therapy for human influenza virus infection.
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影响因子：
5.4
作者：
Arrighi, JF;Pion, M;Piguet, V
通讯作者：
Piguet, V
影响因子：
9.2
作者：
Doi, N;Zenno, S;Saigo, K
通讯作者：
Saigo, K
DOI：
10.1073/pnas.83.17.6282
发表时间：
1986-09-01
影响因子：
11.1
作者：
BEATON, AR;KRUG, RM
通讯作者：
KRUG, RM
影响因子：
--
作者：
Fish RJ;Kruithof EK
通讯作者：
Kruithof EK
影响因子：
64.8
作者：
Bernstein, E;Caudy, AA;Hannon, GJ
通讯作者：
Hannon, GJ