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

核心C -项目总结/摘要 核心C为研究项目1（RP 01）、RP 02、RP 03和核心的高生物防护工作提供支持 B。通过核心A确定工作的优先次序。核心人员带来分子生物学、细胞生物学、 以及埃博拉和马尔堡病毒的动物疾病模型。核心维持病毒库存， 重组病毒，提供来自感染细胞的细胞裂解物和RNA，并确保它们不具有感染性。 材料，评估宿主因子丢失或过度表达对感染的影响，并进行转基因研究。 啮齿动物这些核心活动提供所需的蛋白质材料和亚细胞结构的理解 RP 01病毒感染后制备，用于评价病毒蛋白功能和基因组茎环 RP 02的转录控制结构，并提供感染动力学和宿主病毒的详细分析 RP 03的联合体。为了直接解决宿主因素对病毒蛋白产生的影响，病毒RNA 转录和亚结构形成，我们已经开发了一种新的蛋白质-抗体+ vRNA-FISH 基于染色技术，通过自动显微图像分析进行定量评估。这 预计这种方法将为每个项目和核心项目B的需求提供重要信息。核心C 通过提供纯化的病毒裂解物，与核心B在新型抗体的生产和评价方面密切合作 并进行感染细胞染色的显微镜评价。该项目的特点是提供 BSL 4的高水平分子生物学能力将用于生产预计具有以下功能的重组病毒 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用于 评估在控制疾病方面的作用。在这样做时，Core C提供了必要的过程来评估主机 从细胞到疾病的因子作用，使得能够识别治疗开发的潜在靶点。

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.