West/Midwest Consortium for High-Resolution Cryo Electron Microscopy

西部/中西部高分辨率冷冻电子显微镜联盟

• 批准号: 10019566
• 负责人: Mavis Agbandje-Mckenna
• 金额: $ 53.9万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-11 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10019566
• 关键词: Alabama; Biochemistry; Biological; Biology; Biomedical Research; California; Characteristics; Communities; Core Facility; Cryoelectron Microscopy; Data; Detection; Development; Drops; Drug Design; Electron Microscope; Electron Microscopy Facility; Electrons; Evaluation; Facility Accesses; Florida; Funding; Goals; Heavy Metals; Image; Infrastructure; Institutes; Kansas; Laboratories; Liquid substance; Los Angeles; Macromolecular Complexes; Medical; Microbiology; Microscope; Midwestern United States; Modeling; Nitrogen; Organelles; Performance; Phase; Records; Research; Research Personnel; Reservations; Resolution; Resources; Sampling; Site; Speed; Stains; Structure; System; Technology; Texas; Therapeutic Intervention; Time; Titan; Training; Translational Research; Transmission Electron Microscopy; United States National Institutes of Health; Universities; Vaccines; Washington; base; data acquisition; design; detector; experience; imaging detection; improved; instrument; member; microscopic imaging; movie; nanometer resolution; nanosystems; new therapeutic target; novel therapeutics; operation; public health relevance; remote control; virology

 DESCRIPTION (provided by applicant): This application seeks funding to establish the West/Midwest Consortium for High-Resolution Cryo Electron Microscopy (cryoEM). The goal of the consortium is to offer 19 cryoEM users from 10 regional institutes free access to a high-end cryoEM facility with proven high-resolution capabilities located in the California NanoSystems Institute (CNSI) at University of California, Los Angeles (UCLA). UCLA will act as host institute and provide investigators in these cryoEM laboratories access to its highly productive Titan Krios cryo electron microscope recently upgraded with a Volta phase plate, a Gatan imaging filter (GIF), and pre- and post-GIF direct electron detectors. A highly experienced staff with proven records of cryoEM-derived atomic structures will provide on-site assistance to collect data and provide streamlined "movie" pre-processing for our consortium user laboratories. The critical need for this consortium for recording atomic-resolution cryoEM images of a broad range of bio-medically significant macromolecular complexes is justified by the immediate benefits to the 19 consortium users. Of particular note, we have demonstrated that, once trained by our staff, users no longer have to be present at the microscope and can remotely control the microscope and perform high-resolution cryoEM imaging after our staff have aligned the instruments for top performance, thus significantly improving accessibility to the high-end instrument. A three-member supervisory committee representing non-consortium users, regional consortium users and the UCLA facility staff will provide evaluation and approval of user projects, adding and dropping users. Under our typical operation, a user group with approved projects requests research instrument time with an existing professionally-designed online reservation system, sends cryoEM grids by overnight courier (such as FedEx) in a liquid nitrogen-cooled dry shipper, and, after their grids are loaded by our staff, image their sample remotely for 3-7 days on average, and up to 2-3 weeks if necessary. The 10 institutes are concentrated on the West coast (University of Washington in Seattle, University of California at San Diego, Scripps) and Midwest (Case Western Reserve University, Purdue University, The University of Kansas, University of Alabama at Birmingham) except for three (University of Texas at El Paso, University of Texas Medical Branch in Galveston, and University of Florida at Gainesville). With the exception of three new investigators, all these users are funded by the NIH or other federal agencies to pursue biomedical research, ranging from biology, biochemistry, virology and microbiology. The establishment of this consortium will immediately empower our users with high-resolution cryoEM capabilities for a broad range of biological samples, enabling them to understand mechanisms of action and identify new targets for the development of new therapeutics.

 描述（由申请人提供）：本申请寻求资金建立西部/中西部高分辨率冷冻电子显微镜联盟（cryoEM）。该联盟的目标是为来自10个地区研究所的19名cryoEM用户免费提供位于洛杉矶加州大学（UCLA）加州纳米系统研究所（CNSI）的高端cryoEM设施，该设施具有经过验证的高分辨率能力。加州大学洛杉矶分校将作为主办机构，并为这些cryoEM实验室的研究人员提供其高效的Titan Krios低温电子显微镜，该显微镜最近升级了Volta相位板，Gatan成像滤波器（GIF）以及GIF前后的直接电子探测器。一个经验丰富的工作人员与cryoEM衍生的原子结构的证明记录将提供现场协助收集数据，并提供精简的“电影”预处理我们的联盟用户实验室。该财团记录原子分辨率cryoEM图像的广泛的生物医学显着的大分子复合物的迫切需要是合理的直接利益的19个财团的用户。特别值得注意的是，我们已经证明，一旦经过我们的工作人员培训，用户不再需要在显微镜前，并且可以远程控制显微镜，并在我们的工作人员将仪器对准以获得最佳性能后执行高分辨率cryoEM成像，从而显着提高高端仪器的可访问性。一个由三名成员组成的监督委员会代表非财团用户、区域财团用户和加州大学洛杉矶分校设施工作人员，将对用户项目、增加和删除用户进行评估和批准。在我们的典型操作下，具有批准项目的用户组通过现有的专业设计的在线预订系统请求研究仪器时间，通过隔夜快递（如联邦快递）在液氮冷却的干燥托运箱中发送cryoEM网格，并且，在我们的工作人员装载网格后，远程成像他们的样品平均3-7天，如果需要，最多2-3周。这10所研究所集中在西海岸（西雅图的华盛顿大学，加州大学圣地亚哥分校，斯克里普斯）和中西部（凯斯西储大学，普渡大学，堪萨斯大学，亚拉巴马大学伯明翰分校），除了三所（德克萨斯大学埃尔帕索分校，德克萨斯大学加尔维斯顿分校和佛罗里达大学盖恩斯维尔分校）。除了三名新的研究人员外，所有这些用户都由NIH或其他联邦机构资助，从事生物医学研究，包括生物学、生物化学、病毒学和微生物学。该联盟的成立将立即为我们的用户提供针对广泛生物样品的高分辨率cryoEM能力，使他们能够了解作用机制并确定新疗法开发的新靶点。

项目成果

Structural basis of TRPV5 regulation by physiological and pathophysiological modulators.

• DOI: 10.1016/j.celrep.2022.110737
• 发表时间: 2022-04-26
• 期刊: CELL REPORTS
• 影响因子: 8.8
• 作者: Fluck, Edwin C.;Yazici, Aysenur Torun;Rohacs, Tibor;Moiseenkova-Bell, Vera Y.
• 通讯作者: Moiseenkova-Bell, Vera Y.

Fibril structures of TFG protein mutants validate the identification of TFG as a disease-related amyloid protein by the IMPAcT method.

• DOI: 10.1093/pnasnexus/pgad402
• 发表时间: 2023-12
• 期刊: PNAS NEXUS
• 作者: Rosenberg, Gregory M.;Abskharon, Romany;Boyer, David R.;Ge, Peng;Sawaya, Michael R.;Eisenberg, David S.
• 通讯作者: Eisenberg, David S.

Atomic model of vesicular stomatitis virus and mechanism of assembly.

• DOI: 10.1038/s41467-022-33664-4
• 发表时间: 2022-10-10
• 期刊: Nature communications
• 影响因子: 16.6

Near-Atomic-Resolution Cryo-Electron Microscopy Structures of Cucumber Leaf Spot Virus and Red Clover Necrotic Mosaic Virus: Evolutionary Divergence at the Icosahedral Three-Fold Axes.

黄瓜叶斑病毒和红三叶草坏死花叶病毒的近原子分辨率冷冻电子显微镜结构：二十面体三重轴的进化分歧。

• DOI: 10.1128/jvi.01439-19
• 发表时间: 2020
• 期刊: Journal of virology
• 影响因子: 5.4
• 作者: Sherman,MichaelB;Guenther,Richard;Reade,Ron;Rochon,D'Ann;Sit,Tim;Smith,ThomasJ
• 通讯作者: Smith,ThomasJ

Structure of Tetrahymena telomerase-bound CST with polymerase α-primase.

• DOI: 10.1038/s41586-022-04931-7
• 发表时间: 2022-08
• 期刊: Nature
• 影响因子: 64.8