High Biocontainment (BSL4/ABSL4) core for replication competent virus work

核心C--项目摘要/摘要 核心C为研究项目1(RP01)、RP02、RP03和核心提供高生物遏制工作的支持 B.通过核心A优先开展工作。核心人员带来分子生物学、细胞生物学等方面的独特专业知识 以及针对埃博拉和马尔堡病毒的动物疾病建模。核心维护病毒库存，使 重组病毒，提供受感染细胞的细胞裂解产物和RNA，并确保它们不受感染 材料，评估宿主因子丢失或过表达对感染的影响，并进行转基因工作 啮齿动物。这些核心活动提供了所需的蛋白质物质和对亚细胞结构的理解 病毒感染后制作的RP01，是评估病毒蛋白功能和基因组干环所必需的 结构控制RP02的转录，并提供感染动力学和宿主病毒的详细分析 RP03的联合协会。为了直接解决宿主因素对病毒蛋白生产的影响，病毒RNA 转录和各自形成的亚结构，我们开发了一种新的蛋白质-抗体vRNA-FISH 基于染色技术，通过自动显微镜图像分析进行定量评估。这 预计该方法将为每个项目和核心B的需求提供重要信息。 通过提供纯化的病毒裂解物，与Core B密切合作生产和评估新抗体 以及对感染细胞进行染色的显微镜评估。该项目的突出之处在于提供 BSL4的高级分子生物学能力将用于生产重组病毒，预计具有 改变了复制动力学。此外，通过核心B提供的转基因动物的使用将被用于 评估在控制疾病中的作用。在此过程中，Core C提供了评估主机所需的流程 从细胞到疾病的因子作用，使确定潜在的治疗开发目标成为可能。

Core C – Project Summary/Abstract Core C provides support for high biocontainment work to Research Projects 1 (RP01), RP02, RP03 and Core B. Work is prioritized through Core A. Core personnel bring unique expertise in molecular biology, cell biology and animal disease modeling for Ebola and Marburg viruses. The core maintains virus stocks, makes recombinant viruses, provides cell lysates and RNA from infected cells and ensures they are free of infectious material, evaluates effects of host factor loss or overexpression on infection and performs work with transgenic rodents. These core activities provide needed protein material and an understanding of subcellular structures made after virus infection for RP01, are required for evaluation of virus protein function and genome stem loop structures in controlling transcription for RP02, and provide detailed analysis of infection kinetics and host-virus co-association for RP03. To directly address impact of host factors on virus protein production, virus RNA transcription and the substructures formed by each, we have developed a novel protein-antibody + vRNA-FISH based staining technique that is quantitatively assessed by automated microscopy image analysis. This approach is expected to provide important information for the needs of each project and Core B. Core C also works closely with Core B in production and evaluation of novel antibodies by providing purified virus lysates and performing microscopy evaluation of staining of infected cells. The project distinguishes itself in providing high-level molecular biology ability at BSL4 that will be used to produce recombinant viruses predicted to have altered replication kinetics. Furthermore, use of transgenic animals, provided through Core B, will be used to evaluate roles in controlling disease. In so doing, Core C provides the necessary processes to evaluate host factor roles from cells through to disease that enables identification of potential targets for therapy development.