SARS Coronavirus Reverse Genetics and Pathogenesis

SARS冠状病毒反向遗传学和发病机制

• 批准号: 6908934
• 负责人: Paul Scott Masters
• 金额: $ 27.4万
• 依托单位: WADSWORTH CENTER
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-07-01 至 2007-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6908934
• 关键词: RNA biosynthesis; SARS virus; disease /disorder model; gene expression; genetically modified animals; hepatitis virus; host organism interaction; laboratory mouse; method development; model design /development; mutant; phenotype; polymerase chain reaction; recombinant virus; severe acute respiratory syndrome; tissue /cell culture; virus RNA; virus genetics; virus infection mechanism; virus protein

DESCRIPTION (provided by applicant): Coronaviruses are a family of enveloped, single-stranded, positive-sense RNA viruses. The genomes of coronaviruses are the largest among all the RNA viruses, which has made their genetic manipulation a formidable problem. Our laboratory developed the first reverse genetic system for coronaviruses, called targeted RNA recombination. This system has been used to answer fundamental questions about viral protein structure and function, host species specificity, virion assembly, and the unusually complex mechanism of coronavirus genome RNA synthesis. Targeted RNA recombination is a powerful and versatile method that, in principle, should be applicable to all species of coronaviruses. The major object of this application is to develop and apply this system to the coronavirus that is the causative agent for Severe Acute Respiratory Syndrome (SARS-CoV). To accomplish this, we will construct an interspecies chimeric coronavirus that will have gained the ability to infect mouse cells. This virus, designated mSARS-CoV, will serve as the cornerstone for a host-range-based selection system for SARS-CoV reverse genetics. The tissue culture properties of mSARS-CoV and mutants of mSARS-CoV will be explored to gain insights into the potential roles of the eight unique genes in SARS-CoV that do not appear in other coronaviruses. The disease caused by these constructed viruses in the mouse host will be characterized and compared to that caused by the well-studied mouse coronavirus (MHV). Additionally, this genetic system will be used to answer basic questions about the components of SARS-CoV virion assembly and RNA synthesis and to create a mouse virus surrogate of SARS-CoV whose virion proteins are completely derived from MHV. The proposed work is expected to produce significant new information upon which more detailed future studies will be based. It will also generate valuable tools to test antiviral drugs, reveal targets for such drugs, and provide a means to manipulate SARS-CoV for vaccine design.

描述（由申请人提供）：冠状病毒是一类有包膜、单链、正义 RNA 病毒。冠状病毒的基因组是所有RNA病毒中最大的，这使得它们的基因操作成为一个艰巨的问题。我们的实验室开发了第一个冠状病毒反向遗传系统，称为靶向RNA重组。该系统已被用来回答有关病毒蛋白结构和功能、宿主物种特异性、病毒体组装以及冠状病毒基因组 RNA 合成异常复杂机制的基本问题。靶向 RNA 重组是一种强大且通用的方法，原则上应适用于所有冠状病毒物种。该应用的主要目标是开发该系统并将其应用于作为严重急性呼吸系统综合症（SARS-CoV）病原体的冠状病毒。为了实现这一目标，我们将构建一种具有感染小鼠细胞能力的种间嵌合冠状病毒。这种病毒被命名为 mSARS-CoV，将作为 SARS-CoV 反向遗传学基于宿主范围的选择系统的基石。将探索 mSARS-CoV 和 mSARS-CoV 突变体的组织培养特性，以深入了解 SARS-CoV 中其他冠状病毒中不存在的八个独特基因的潜在作用。这些构建的病毒在小鼠宿主中引起的疾病将被表征，并与经过充分研究的小鼠冠状病毒（MHV）引起的疾病进行比较。此外，该遗传系统将用于回答有关 SARS-CoV 病毒体组装和 RNA 合成成分的基本问题，并创建 SARS-CoV 的小鼠病毒替代品，其病毒体蛋白完全来源于 MHV。拟议的工作预计将产生重要的新信息，未来更详细的研究将以此为基础。它还将产生有价值的工具来测试抗病毒药物，揭示此类药物的靶点，并提供一种操纵 SARS-CoV 进行疫苗设计的方法。