A Mid-Level 200kV Instrument for Single-Particle cryoEM

用于单粒子冷冻电镜的中级 200kV 仪器

• 批准号: 10436739
• 负责人: Z Hong ZHOU
• 金额: $ 200万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-20 至 2024-06-19
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10436739
• 关键词: 14 year old; Architecture; Biological; Biomedical Research; California; Communities; Complex; Cryo-electron tomography; Cryoelectron Microscopy; Data Set; Data Sources; Electrons; Engineering; Ensure; Faculty; Funding; Goals; Image; Imaging Techniques; Institutes; Los Angeles; Micro Electron Diffraction; Natural Sciences; Neurodegenerative Disorders; Nucleoproteins; Penetration; Research; Research Project Grants; Resolution; Resources; Spectrometry; Structure; System; Telomerase; Time; Titan; United States National Institutes of Health; Universities; X-Ray Crystallography; age related; bactericide; base; biological research; college; imaging capabilities; imaging facilities; instrument; interest; medical schools; member; method development; nanobiology; nanomachine; nanoscale; nanosystems; particle; structural biology

Project Summary The goal of the California NanoSystems Institute (CNSI) at the University of California at Los Angeles (UCLA) is to enable cutting-edge nanobiology and biomedical research. As a key part of this goal, the Electron Imaging Center for Nanomachines (EICN) was established in 2007 to enable necessary advanced imaging techniques for visualizing and understanding the mechanisms of macromolecular machineries at the nanometer scale. Funding from a previous NIH S10 to EICN was instrumental to the purchase (2006) and installation (2007) of the world’s first working Titan Krios instrument, which contributed to the ‘cryoEM revolution’ of the last decade. This application seeks funding to purchase a Glacios, a mid-level 200 kV instrument with demonstrated high- resolution single-particle cryoEM imaging capability, to bridge the existing sets of entry-level and high-end Titan Krios instruments at the EICN at UCLA. The critical need for this instrument is justified as follows: First, the new instrument will meet the pressing need for single-particle cryoEM imaging in a broad range of federally funded biomedical research projects by 24 UCLA labs, including telomerase and spliceosomal nucleoprotein complexes; trans-membrane channels, transporters, bactericidal machineries and secretion systems; and complexes involved in neuro-degenerative diseases. Second, the existing 14-year old Titan Krios has developed age-related instability issues; additionally, 50% of its accessible time is reserved for an NIH U24 user consortium, while the other 50% is used to meet the quickly growing need of cryo-electron tomography (cryoET) users at UCLA, thanks to its high penetration power of 300 keV electrons and BioQuantum K3 Imaging filter. UCLA-based research would be greatly enhanced by the acquisition of this mid-level cryoEM for atomic structure determination by the single-particle cryoEM approach described in the application. The proposed instrument will enjoy strong institutional support thus ensuring lasting impact. High-resolution electron imaging has become an integral part of CNSI’s highly successful research resources and of the very strong structural biology research community at UCLA. The identification of tens of users with active federal funding shows that there is major interest across multiple departments/institutes among the colleges of natural sciences and engineering, as well as the UCLA medical school, which may greatly benefit from acquiring this mid-level cryoEM instrument with demonstrated high-resolution cryoEM capability. This instrument, together with existing micro-electron diffraction (microED), X-ray crystallography and NMR spectrometry, will provide a robust resource for faculty members who are eager to expand the scope of their current biomedical and biological research projects to include cryoEM. The new instrument will also meet a critical need for huge data sets for pushing the envelope of cryoEM for the cryoID method development. The diverse biological structures to be studied, and their highly varied architectures, offer a fertile data source for the scientific community for method development pursued by UCLA and the general electron imaging community at large.

项目概要 加州大学洛杉矶分校 (UCLA) 加州纳米系统研究所 (CNSI) 的目标 旨在实现尖端的纳米生物学和生物医学研究。作为这一目标的关键部分，电子成像 纳米机器中心 (EICN) 成立于 2007 年，旨在实现必要的先进成像技术 用于在纳米尺度上可视化和理解大分子机械的机制。 以前的 NIH S10 向 EICN 提供的资金有助于购买（2006 年）和安装（2007 年） 世界上第一台可用的 Titan Krios 仪器，为过去十年的“冷冻电镜革命”做出了贡献。 本申请寻求资金购买 Glacios，这是一款中级 200 kV 仪器，具有高可靠性 分辨率单粒子冷冻电镜成像能力，连接现有的入门级和高端套 加州大学洛杉矶分校 EICN 的 Titan Krios 仪器。对该仪器的迫切需求理由如下： 首先，新仪器将满足广泛领域对单颗粒冷冻电镜成像的迫切需求 加州大学洛杉矶分校 24 个实验室的联邦资助生物医学研究项目，包括端粒酶和剪接体 核蛋白复合物；跨膜通道、转运蛋白、杀菌机制和分泌 系统；以及与神经退行性疾病有关的复合物。二、现有14岁泰坦克里奥斯 出现了与年龄相关的不稳定问题；此外，50% 的可用时间预留给 NIH U24 用户联盟，另外50%用于满足快速增长的冷冻电子断层扫描需求 加州大学洛杉矶分校 (cryoET) 用户，得益于其 300 keV 电子的高穿透力和 BioQuantum K3 成像滤镜。通过购买这种中级冷冻电镜，加州大学洛杉矶分校的研究将得到极大加强 通过应用中描述的单粒子冷冻电镜方法测定原子结构。 拟议文书将获得强有力的机构支持，从而确保产生持久影响。高分辨率 电子成像已成为 CNSI 非常成功的研究资源和非常重要的研究资源的一个组成部分。 加州大学洛杉矶分校强大的结构生物学研究社区。数十名活跃联邦用户的身份识别 资金表明，自然学院的多个部门/研究所都表现出浓厚的兴趣 科学与工程，以及加州大学洛杉矶分校医学院，可能会从获得这个项目中受益匪浅 具有高分辨率冷冻电镜功能的中级冷冻电镜仪器。这个乐器，一起 结合现有的微电子衍射（microED）、X射线晶体学和核磁共振波谱测定，将提供 为渴望扩大当前生物医学和生物医学领域范围的教师提供强大的资源 生物研究项目包括冷冻电镜。新仪器还将满足对海量数据的迫切需求 为 CryoID 方法开发突破 CryoEM 的极限。多样化的生物结构 有待研究，以及它们高度多样化的架构，为科学界提供了丰富的数据源 加州大学洛杉矶分校和整个电子成像界所追求的方法开发。