NIEHS Cryo-EM Core Facility

NIEHS Cryo-EM核心继续在该地区领导部署和促进采用低温电子显微镜技术。2022财年在高影响力期刊上发表的十几篇同行评议文章就是一个明显的迹象。CORE在应对新冠肺炎大流行的内部应对中发挥了重要作用，有四份同行评议的出版物描述了对抗SARS-CoV-2的潜在药理靶点或试剂(Hong等人。PNAS 2022，Fu等人，PLOS One 2022，Frazier等人。核酸研究报告2022，Frazier等人。DNA Res.2021和Pillon等人。纳特。通信。)。许多合作为理解疾病和治疗的结构基础做出了重要贡献。这些在核心解决的疾病相关结构涉及线粒体遗传病(Riccio等人)。PNAS)、帕金森与痴呆(Dasari等人，《生物化学》2022)、抗癌药物输送系统(Saha等人Nano Lett.2022)，抗菌素耐药性(Travis等人纳特。交警。2022年)。其他结构参与基本的生物过程，如疼痛、寒冷和热感觉(权纳特结构摩尔生物)。2021)，代谢途径(Klemm等人。J Biol Chem.)，以及对细胞应激的响应(Simos等人细胞代表2022，Kumar等人。自然2022)。 核心得益于与分子显微镜小组在开发方法方面的密切合作，以增强我们的技术能力。这些措施包括部署用于样品沉积和时间分辨结构确定的内部工程机器人设备，改进和加速数据收集方法，以及在线处理和评估数据的软件。今年，我们通过部署与NIA、NIBIB、NIDA和NCATS合作购买的300keV冷冻电子显微镜，扩大了我们的能力。这一最先进的仪器将通过NIH Cryo-EM联盟向合作研究所和IRP的其他机构提供单粒子Cryo-EM的高通量和高分辨率数据收集。我们还正在完成部署低温聚焦离子束扫描电子显微镜(CRYO-FIBSEM)。这台仪器将使我们能够深入细胞，在原位解决大分子络合物的结构。

The NIEHS Cryo-EM Core continues to lead the deployment and to promote the adoption of techniques in cryo-electron microscopy in the region. Over a dozen peer reviewed articles in high impact journals during FY2022 are a clear indication. The Core has played an important role in the intramural response to the COVID-19 pandemic with four peer reviewed publications describing potential pharmacological targets or reagents against SARS-CoV-2 (Hong et al. PNAS 2022, Fu et al, PloS One 2022, Frazier et al. Nucleic Acids Res. 2022, Frazier et al. Nucleic Acids Res. 2021 and Pillon et al. Nat. Commun.). Many collaborations have led to important contributions to the understanding of structural underpinnings of disease and therapy. These disease relevant structures solved at the Core involve mitochondrial genetic disease (Riccio et al. PNAS), Parkinson and dementia ( Dasari et al., Biochemistry 2022), anticancer drug delivery systems (Saha et al Nano Lett. 2022), antibacterial resistance (Travis et al. Nat. Commun. 2022). Other structures are involved in fundamental biological processes such as pain, cold and heat sensing (Kwon Nat Struct Mol Biol. 2021), metabolic pathways (Klemm et al. J Biol Chem.), and responses to cellular stress (Simoes et al. Cell Rep. 2022, Kumar et al. Nature 2022). The Core benefits from a close collaboration with the Molecular Microscopy Group on the development of methods that enhance our technical capabilities. These include deployment of in house engineered robotic equipment for sample deposition and time resolved structure determination, improved and accelerated methods for data collection, and software for in-line processing and assessment of data. This year we expanded our capabilities by deploying a 300 KeV cryo-electron microscope acquired in collaboration with NIA, NIBIB, NIDA and NCATs. This state-of-the-art instrument will provide access to high-throughput and high-resolution data collection for single particle cryo-EM to the collaborating institutes and to others at the IRP via the NIH Cryo-EM Consortium. We are also completing the deployment of a cryogenic focused ion beam scanning electron microscope (cryo-FIBSEM). This instrument will allow us to delve into the cell to solve the structure of macromolecular complexes in situ.