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AN ORGANISMAL APPROACH TO VIRAL HOST FACTOR DISCOVERY

发现病毒宿主因子的有机方法

基本信息

批准号：
7563272
负责人：
RICHARD W HARDY
金额：
$ 18.5万
依托单位：
TRUSTEES OF INDIANA UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-02-15 至 2011-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7563272
关键词：
Alphavirus Antiviral Agents Antiviral Therapy Biochemical Bioinformatics Cells Cellular Structures Chikungunya virus Chromosome Deletion Classification Code Collaborations Complex Culicidae Development Developmental Process Disease Drosophila genome Drosophila genus Drosophila melanogaster Eastern Equine Encephalitis Virus Engineering Environment Epidemic Firefly Luciferases Gene Expression Genes Genetic Genetic Models Genetic Transcription Genome Genomics Goals Human Indiana Individual Infection Integration Host Factors Lead Livestock Location Maps Messenger RNA Modification Molecular Molecular Genetics Molecular Target Monitor Morbidity - disease rate Morphogenesis Mutation Organism Pattern Physiological Poly A Population Proteins Public Health RNA RNA Sequences Replicon Reporter Reporter Genes Research Resources Structural Protein System Testing Time Tissues Universities Vaccines Venezuelan Equine Encephalitis Virus Viral Viral Genes Viral Genome Viral Proteins Virion Virus Virus Replication Work design fly genetic manipulation mortality novel pathogen programs public health relevance therapy development tool virus host interaction

项目摘要

DESCRIPTION (provided by applicant): Alphaviruses are pathogens of humans and livestock with worldwide distribution. The classification of some species as select agents in conjunction with their impact on public health makes understanding their interaction with the host and development of interventions a high priority. A major impediment to understanding the host cell components with which the virus interacts has been the inability to genetically manipulate the host cell/organism. Drosophila melanogaster represents an excellent experimental system for elucidating the genetic, molecular, and biochemical mechanisms underlying numerous physiological and developmental processes. The proposed research aims to exploit the power of Drosophila genetics for the discovery of host factors required for alphavirus genome replication. The basic concept for the research is straightforward; an alphavirus replicon sequence encoding a reporter/selectable marker will be cloned into the Drosophila genome under the control of the UAS/GAL4 system. Flies containing the replicon sequence will be crossed with a GAL4 expressing line to direct primary transcription of the replicon RNA in F1 progeny. If the cellular environment in which the replicon is expressed is permissive for virus genome replication then replicon amplification and reporter gene expression will occur. This approach will allow (i) the identification of tissues throughout the fly capable of supporting virus replication; (ii) genetic identification of host factors required for viral genome replication. The research outlined in the proposal is not complex, involving relatively simple manipulations of the Drosophila and alphaviral genomes; however the potential payoff is enormous. This simple but powerful approach will lead to the identification of dozens if not hundreds of host factors required for viral genome replication, and significantly advance our understanding of the virus-host interaction. PUBLIC HEALTH RELEVANCE: Identifying the ways in which a virus interacts with it's host is crucial to understanding the underlying cause of disease and developing antiviral drugs. The proposed research aims to determine host factors required for alphavirus (mosquito-borne pathogens) infection using a model genetic system.

描述（由申请人提供）：甲病毒是人类和牲畜的病原体，在世界范围内分布。将某些物种分类为选定的媒介，并结合它们对公共健康的影响，使了解它们与宿主的相互作用和制定干预措施成为当务之急。了解与病毒相互作用的宿主细胞成分的一个主要障碍是无法对宿主细胞/生物体进行遗传操作。黑腹果蝇代表了一个出色的实验系统，用于阐明众多生理和发育过程背后的遗传、分子和生化机制。拟议的研究旨在利用果蝇遗传学的力量来发现甲病毒基因组复制所需的宿主因子。这项研究的基本概念很简单；编码报告/选择标记的甲病毒复制子序列将在 UAS/GAL4 系统的控制下克隆到果蝇基因组中。含有复制子序列的果蝇将与 GAL4 表达系杂交，以指导 F1 后代中复制子 RNA 的初级转录。如果表达复制子的细胞环境允许病毒基因组复制，则复制子扩增和报告基因表达将会发生。这种方法将允许（i）识别整个苍蝇中能够支持病毒复制的组织； (ii) 病毒基因组复制所需宿主因子的遗传鉴定。提案中概述的研究并不复杂，涉及对果蝇和甲病毒基因组的相对简单的操作；然而，潜在的回报是巨大的。这种简单但强大的方法将导致病毒基因组复制所需的数十甚至数百个宿主因子的鉴定，并显着增进我们对病毒与宿主相互作用的理解。公共卫生相关性：确定病毒与其宿主相互作用的方式对于了解疾病的根本原因和开发抗病毒药物至关重要。拟议的研究旨在使用模型遗传系统确定甲病毒（蚊媒病原体）感染所需的宿主因素。