Drosophila RNAi screen identifies host genes important for influenza virus replication.
Drosophila RNAi screen identifies host genes important for influenza virus replication.
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DOI：
10.1038/nature07151
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发表时间：
2008-08-14
期刊：
影响因子：
64.8
通讯作者：
Kawaoka, Yoshihiro
中科院分区：
文献类型：
--
作者：
Hao, Linhui;Sakurai, Akira;Watanabe, Tokiko;Sorensen, Ericka;Nidom, Chairul A.;Newton, Michael A.;Ahlquist, Paul;Kawaoka, Yoshihiro
所有病毒都依赖宿主细胞蛋白及其相关机制来完成病毒生命周期。识别参与病毒复制每一步的宿主分子可能为抗病毒治疗提供有价值的新靶点，但这一目标可能需要数十年的传统正向基因筛查方法和哺乳动物细胞培养才能实现。在这里，我们描述了一种在果蝇中的新的全基因组RNA干扰(RNAi)筛查，它可以用于识别对流感病毒复制重要的宿主基因。在修改流感病毒以允许感染果蝇细胞和检测流感病毒基因表达后，我们测试了针对13,071个基因(90%的果蝇基因组)的RNAi文库，鉴定出超过100个基因在果蝇细胞中的抑制显著抑制或刺激了流感病毒衍生载体的报告基因(Renilla荧光素酶)的表达。这些发现与哺乳动物细胞的流感病毒感染有关，上面确定的果蝇基因的一个子集也说明了这一点。也就是说，ATP6V0d1、COX6A1和Nxf1的人类同源物被证明在H5N1和H1N1甲型流感病毒的复制中具有关键功能，但不能在HEK 293细胞中复制水泡性口炎病毒或牛痘病毒。因此，我们已经证明了在果蝇中使用全基因组RNAi筛选来识别流感病毒复制所需的先前未识别的宿主蛋白的可行性。这可能会加速开发用于化学预防和治疗的新型抗病毒药物，鉴于快速开发有效的大流行性流感疫苗的障碍以及可能出现对现有药物具有抗药性的菌株，这些药物是迫切需要的。
All viruses rely on host cell proteins and their associated mechanisms to complete the viral life cycle. Identifying the host molecules that participate in each step of virus replication could provide valuable new targets for antiviral therapy, but this goal may take several decades to achieve with conventional forward genetic screening methods and mammalian cell cultures. Here we describe a novel genome-wide RNA interference (RNAi) screen in Drosophila that can be used to identify host genes important for influenza virus replication. After modifying influenza virus to allow infection of Drosophila cells and detection of influenza virus gene expression, we tested an RNAi library against 13,071 genes (90% of the Drosophila genome), identifying over 100 whose suppression in Drosophila cells significantly inhibited or stimulated reporter gene (Renilla luciferase) expression from an influenza virus-derived vector. The relevance of these findings to influenza virus infection of mammalian cells is illustrated for a subset of the Drosophila genes identified above. That is, the human homologues of ATP6V0D1, COX6A1 and NXF1 are shown to have key functions in the replication of H5N1 and H1N1 influenza A viruses, but not vesicular stomatitis virus or vaccinia virus, in HEK 293 cells. Thus, we have demonstrated the feasibility of using genome-wide RNAi screens in Drosophila to identify previously unrecognized host proteins that are required for influenza virus replication. This could accelerate the development of new classes of antiviral drugs for chemoprophylaxis and treatment, which are urgently needed given the obstacles to rapid development of an effective vaccine against pandemic influenza and the likely emergence of strains resistant to available drugs.
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影响因子：
3.8
作者：
PEREZ, L;CARRASCO, L
通讯作者：
CARRASCO, L
影响因子：
5.4
作者：
WYERS, F;RICHARDMOLARD, C;DEZELEE, S
通讯作者：
DEZELEE, S
DOI：
10.1073/pnas.0610977104
发表时间：
2007-02-06
影响因子：
11.1
作者：
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通讯作者：
Fontoura, Beatriz M. A.
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3.7
作者：
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通讯作者：
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影响因子：
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作者：
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通讯作者：
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