Antiviral effects against influenza A virus infection by a short hairpin RNA targeting the non-coding terminal region of the viral nucleoprotein gene
Antiviral effects against influenza A virus infection by a short hairpin RNA targeting the non-coding terminal region of the viral nucleoprotein gene
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DOI：
10.1292/jvms.18-0436
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发表时间：
2019-03-01
影响因子：
1.2
通讯作者：
Ono, Etsuro
中科院分区：
文献类型：
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作者：
Fujimoto, Yoshikazu;Kyogoku, Kenji;Ono, Etsuro
RNA干扰（RNAi）可以以基因特异性方式抑制甲型流感病毒（IAV）感染。在这项研究中，我们构建了一个表达短发夹 RNA (shRNA) 的转基因，该转基因靶向编码核蛋白 (NP) 的 IAV RNA 基因的非编码区域。为了研究 shRNA 的抗病毒作用，我们用该转基因培育了两个转基因小鼠品系。不幸的是，转基因和非转基因同窝仔猪的 IAV 抗性没有明显差异。为了进一步研究 shRNA 的抗病毒作用，我们从转基因和非转基因小鼠中制备了小鼠胚胎成纤维细胞 (MEF)。在使用这些 MEF 的实验感染中，与非转基因 MEF 相比，转基因 MEF 中小鼠适应的 IAV 毒株 A/Puerto Rico/8/1934 (PR8) 的病毒产生通过在感染早期阶段病毒 RNA 基因转录的下调而受到抑制。这些结果表明，shRNA 的表达能够赋予 MEF 抗 IAV 的抗病毒特性，尽管效果有限。我们的研究结果表明，针对 NP 病毒 RNA (vRNA) 非编码区的 shRNA 可能是开发抗流感家禽的支持工具。
RNA interference (RNAi) can inhibit Influenza A virus (IAV) infection in a gene-specific manner. In this study, we constructed a transgene expressing a short hairpin RNA (shRNA) that targets the noncoding region of the IAV RNA gene encoding nucleoprotein (NP). To investigate the antiviral effects of the shRNA, we generated two transgenic mouse lines with this transgene. Unfortunately, there was no apparent difference in IAV resistance between transgenic and non-transgenic littermates. To further investigate the antiviral effects of the shRNA, we prepared mouse embryonic fibroblasts (MEFs) from transgenic and non-transgenic mice. In experimental infections using these MEFs, virus production of mouse-adapted IAV strain A/Puerto Rico/8/1934 (PR8) in the transgenic MEFs was suppressed by means of the down-regulation of the viral RNA gene transcription in the early stages of infection in comparison with non-transgenic MEFs. These results indicated that expression of the shRNA was able to confer antiviral properties against IAVs to MEFs, although the effects were limited. Our findings suggest that the shRNA targeting the noncoding region of the viral RNA (vRNA) of NP might be a supporting tool in developing influenza-resistant poultry.