Revolutionizing virus-host interaction studies with non-canonical small RNAs

描述（由申请方提供）：甲型流感病毒（IAV）是全球健康的重大负担。这项工作提出了产生IAV毒株的文库，每个毒株编码靶向已知抗病毒宿主基因的siRNA，目的是在体内感染的背景下定义宿主因子的效力。在这些研究中，我们将使用一种小鼠适应的甲型流感病毒，该病毒缺乏全功能的NS1蛋白，从而使病毒显著减毒（mIAV）。该提案的具体目的是（1）工程化基于mIAV的siRNA文库，（2）进行体内RNAi筛选以鉴定IAV复制的宿主决定簇（3）表征鉴定的宿主限制因子。为了完成这项工作，将siRNA文库引入我们的miAV中，并单独拯救病毒。这些病毒被用来感染小鼠，以确定哪些病毒在种群中占主导地位。在初步筛选中鉴定了几种已知的抗病毒因子，包括Trim21，Tlr7， RNaseL、Tbk1和Irf1。此外，还鉴定了两种具有未知抗病毒活性的因子（Zfp182和Mapk 8），目前正在对其进行表征。

DESCRIPTION (provided by applicant): Influenza A virus (IAV) represents a significant burden on global health. This work proposes to generate a library of IAV strains each encoding a siRNA targeting known antiviral host genes with the goal of defining the potency of host factors in the context of an in vivo infection. For these studies, we will use a mouse-adapted influenza A virus that lacks a fully functional NS1 protein, thereby rendering the virus significantly attenuated (mIAV). The specific aims of this proposal are to (1) engineer a mIAV-based siRNA library, (2) perform an in vivo RNAi screen to identify host determinants of IAV replication (3) to characterize the identified host restriction factors. To complete this work, a library of siRNAs was introduced into our mIAV, and the viruses were rescued individually. The viruses were used to infect mice to determine which viruses dominated in the population. Several known antiviral factors were identified in a preliminary screen that includes Trim21, Tlr7, RnaseL, Tbk1 and Irf1. In addition, two factors (Zfp182 and Mapk8) with unknown antiviral activity were identified which are currently being characterized.