Center for Structural Biology of HIV RNA

核心3。成像 总结 CRNA Imaging Core的目的是为CRNA研究者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。芯会 与CRNA研究人员合作，使用高分辨率光学显微镜定位和跟踪HIV-1 RNA， 蛋白质复合物在其细胞背景下，并使用单粒子冷冻电子显微镜（冷冻EM）， 确定RNA、RNA蛋白复合物和HIV相关蛋白复合物的结构。核心会起作用的 通过整合活细胞和超分辨率光， 显微镜的方法，促进了快速改进的方法，在冷冻电子断层扫描（冷冻ET）。的 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。 细胞背景核心实验室由两名PI共同领导。密歇根大学的组成部分侧重于 cryo-EM，由PI Melanie Ohi执导。威斯康星州大学的实验室由PI内森·谢勒管理， 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为 as a repository仓库for materials材料and protocols协议.核心团队将专注于三个目标，包括利用单一 确定RNA、RNA-蛋白质复合物和HIV-1相关蛋白质结构的粒子冷冻EM方法 复杂;开发有效的“样品到结构”管道，以有效分析样品， 核心的成像能力从50毫米到3毫米以下的分辨率;并朝着使用相关的 光学和电子显微镜（CLEM）和冷冻ET检测和确定细胞内的结构。支持 根据其训练使命，核心部队还将为陆军指挥官培训学员制定创新的教育计划， 成为这些方法和技术的独立实践者。

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.