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

利用非经典小 RNA 彻底改变病毒与宿主相互作用的研究

• 批准号: 8926663
• 负责人: Asiel Arturo Benitez
• 金额: $ 3.63万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-01 至 2016-06-24
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8926663
• 关键词: Antiviral Agents; Attenuated; Data; Disease Outcome; Engineering; Ensure; Fellowship; Gene Expression Profile; Gene Silencing; Genes; Goals; Grant; Health; Immune response; Infection; Influenza A virus; Integration Host Factors; Libraries; Methodology; MicroRNAs; Microbe; Molecular Biology; Mus; Orthomyxoviridae; PTPN11 gene; Population; Proteins; RNA Interference; RNA Viruses; RNA library; Recording of previous events; Relative (related person); Shapes; Small Interfering RNA; Small RNA; Specificity; Time; Viral; Virus; Virus Diseases; Virus Replication; Work; base; design; global health; in vivo; novel; public health relevance; research study; response; screening; virus host interaction

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.

说明(申请人提供)：甲型流感病毒(IAV)对全球卫生造成重大负担。这项工作建议建立一个IAV毒株的文库，每个毒株编码一个针对已知抗病毒宿主基因的siRNA，目的是确定宿主因子在体内感染背景下的效力。在这些研究中，我们将使用缺乏全功能NS1蛋白的小鼠适应的甲型流感病毒，从而使病毒显著减毒(MIAV)。这项建议的具体目的是(1)设计一个基于mIAV的siRNA文库，(2)进行体内RNAi筛选，以确定IAV复制的宿主决定因素(3)以表征所识别的宿主限制因素。为了完成这项工作，我们将一个siRNA文库引入到我们的mIAV中，并分别拯救了这些病毒。这些病毒被用来感染老鼠，以确定哪些病毒在种群中占主导地位。在初步筛选中发现了几种已知的抗病毒因子，包括Trim21，Tlr7， RNASEL、TBK1和IRF1。此外，还鉴定了两个目前正在鉴定的具有未知抗病毒活性的因子(Zfp182和Mapk8)。