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

用于复制病毒工作的高生物防护 (BSL4/ABSL4) 核心

基本信息

批准号：
10555054
负责人：
ROBERT A DAVEY
金额：
$ 37.18万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-07-07 至 2028-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10555054
关键词：
129 Mouse Address Amino Acids Animal Disease Models Antibodies Behavior Blood Boston Buffers Cells Cellular biology Clustered Regularly Interspaced Short Palindromic Repeats Cryoelectron Microscopy Disease Disease Outcome Disease model Ebola virus Emerging Communicable Diseases Ensure Evaluation Filoviridae Infections Filovirus GRP94 Genetic Complementation Test Genetic Transcription Genome Histologic Human Resources Image Analysis Immunoprecipitation Impact evaluation Infection Infectious Diseases Research Integration Host Factors Kinetics Knock-out Knockout Mice Liver Marburgvirus Measures Methods Microscopy Modeling Molecular Molecular Biology Mouse Strains Mus Nucleic Acids Outcome Measure Patients Pharmaceutical Preparations Phosphorylation Process Production Proteins Publications RNA Reagent Recombinants Research Project Grants Resources Rodent Role Science Severities Small Interfering RNA Source Stains Structure Subcellular structure Sudan Symptoms System Techniques Testing Training Transcriptional Regulation Transgenic Animals Transgenic Mice Transgenic Organisms Ubiquitination Universities Viral Viral Antigens Viral Proteins Viral load measurement Virion Virus Virus Diseases Western Blotting Work X-Ray Crystallography biosafety level 4 facility design disorder control forest in vivo inhibitor innovation insight knock-down knockout animal knockout gene microscopic imaging mouse model mutant novel novel virus overexpression pathogen pharmacologic programs protein complex protein expression protein function recombinant virus repository reverse genetics small molecule inhibitor stem success targeted treatment therapy development trizol viral RNA viral detection

项目摘要

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.

核心C-项目摘要/摘要 Core C为研究项目1（RP01），RP02，RP03和Core提供了对高生物内在工作的支持 B.通过Core A.核心人员优先考虑工作，为分子生物学，细胞生物学带来独特的经验埃博拉病毒和马堡病毒的动物疾病模型。重组病毒，从感染细胞提供细胞细胞裂解物和RNA，并确保自由的攻击材料，评估宿主因子损失或过表达对感染和性能的影响啮齿动物。评估病毒蛋白功能和基因组茎环的评估需要在病毒感染后制造控制RP02转录的结构，并提供感染动力学和病毒的详细分析 RP03的共同关联。转录和每个形成的子结构，我们开发了一种新型的蛋白质抗体 + vrna-fish 通过自动显微镜图像分析评估定量性染色技术。预计方法有望满足每个项目的需求和核心B。核心核心核心核心核心核心核心核心核心核心核心核心核心核心核心核心核心核心核心核心核心核心核心核心核心核心核心核心核心核心核心核心通过提供纯化的病毒裂解物，与核心箱生产和新型抗体的评估紧密合作并对感染细胞的染色进行显微镜评估。 BSL4的高级分子生物学能力将用于产生预测的重组病毒更改复制动力学。评估控制疾病的作用。从细胞到疾病的因素作用，可以鉴定出潜在的治疗发展。