Center for Structural Biology of HIV RNA

HIV RNA结构生物学中心

• 批准号: 10641982
• 负责人: ALICE TELESNITSKY
• 金额: $ 60.65万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-09 至 2027-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10641982
• 关键词: Biochemical; Cells; Cellular biology; Collaborations; Complex; Cryo-electron tomography; Cryoelectron Microscopy; Development; Education; Electron Microscopy; Goals; HIV; HIV-1; Image; In Vitro; Lead; Methods; Michigan; Mission; Proteins; Protocols documentation; RNA; Research Personnel; Resolution; Resources; Running; Sampling; Scientist; Structure; Techniques; Training; Universities; Virus; Wisconsin; Work; experience; experimental study; high resolution imaging; imaging approach; improved; innovation; light microscopy; particle; programs; protein complex; repository; structural biology; three dimensional structure; tool; ultra high resolution

Core 3. Imaging Summary The purpose of the CRNA Imaging Core is to provide CRNA investigators with access to cutting edge imaging approaches and expertise in both light microscopy and cryo-electron microscopy (cryo-EM). The Core will collaborate with CRNA investigators to use high-resolution light microscopy to localize and track HIV-1 RNA- protein complexes in their cellular context and to use single particle cryo-electron microscopy (cryo-EM) to determine structures of RNAs, RNA-protein complexes, and HIV-related protein complexes. The Core will work to bridge the resolution gap between the two approaches by integrating live cell and superresolution light microscopy approaches, facilitated by the rapidly improving methods in cryo-electron tomography (cryo-ET). The overall goal is to be able to determine 3D structures of HIV RNA-protein complexes both in vitro and in their cellular context. The Core lab is co-led by two PIs. The component at the University of Michigan is focused on cryo-EM and is directed by PI Melanie Ohi. The lab at the University of Wisconsin, run by PI Nathan Sherer, will provide resources and expertise in light microscopy approaches. The University of Michigan Core lab will serve as a repository for materials and protocols. The Core team will focus on three aims that include utilizing single particle cryo-EM approaches to determine structures of RNAs, RNA-protein complexes and HIV-1-related protein complexes; developing effective “sample-to-structure” pipelines for efficient analysis of samples that will leverage the Core’s ability to image at resolutions ranging from ~50 Å to sub-3 Å; and moving towards using correlative light and electron microscopy (CLEM) and cryo-ET to detect and determine structures inside cells. In support of its training mission, the Core will also develop innovative educational programs for CRNA trainees seeking to become independent practitioners of these methods and techniques.

核心 3. 成像 概括 CRNA 成像核心的目的是为 CRNA 研究人员提供最先进的成像技术 光学显微镜和冷冻电子显微镜 (cryo-EM) 的方法和专业知识。核心将 与 CRNA 研究人员合作，使用高分辨率光学显微镜定位和追踪 HIV-1 RNA- 细胞环境中的蛋白质复合物，并使用单粒子冷冻电子显微镜（cryo-EM） 确定 RNA、RNA-蛋白质复合物和 HIV 相关蛋白质复合物的结构。核心将发挥作用 通过集成活细胞和超分辨率光来弥补两种方法之间的分辨率差距 冷冻电子断层扫描（cryo-ET）方法的快速改进促进了显微镜方法的发展。这 总体目标是能够在体外和体内确定 HIV RNA-蛋白质复合物的 3D 结构 细胞环境。核心实验室由两名 PI 共同领导。密歇根大学的该部分重点关注 冷冻电镜由 PI Melanie Ohi 指导。由 PI Nathan Sherer 管理的威斯康星大学实验室将 提供光学显微镜方法的资源和专业知识。密歇根大学核心实验室将服务 作为材料和协议的存储库。核心团队将专注于三个目标，包括利用单一 粒子冷冻电镜方法确定 RNA、RNA-蛋白质复合物和 HIV-1 相关蛋白质的结构 复合体；开发有效的“样品到结构”管道，以有效分析样品，从而利用 Core 能够以约 50 Å 至低于 3 Å 的分辨率成像；并转向使用相关 光学和电子显微镜 (CLEM) 和冷冻电子断层扫描 (cryo-ET) 用于检测和确定细胞内部的结构。支持 根据其培训任务，核心还将为 CRNA 学员开发创新的教育计划，以寻求 成为这些方法和技术的独立实践者。