Opportunities linking omics and structural biology at PNNL: Excelling at Cryo-EM
EMSL Project ID
51710
Abstract
We were recently funded jointly by EMSL and DOE/BER to purchase, install and operate a new Krios G3i cryo-TEM at EMSL. By funding the system jointly, the new microscope would become available to EMSL users 50% and DOE/BER researchers 50% using a resource owner proposal mechanism. The attached project description is meant to act as the umbrella resource owner project for the DOE/BER 50% access. The PI of the project is James Evans and he will act as the resource owner for the duration of the joint operational funding with DOE/BER (currently through FY22). EMSL users will still continue to access this new instrument via the normal EMSL user proposal calls. DOE/BER users will request access to this instrument using the resource owner call. No additional funding from EMSL is being requested as the operational funds from DOE/BER include labor costs for EMSL staff to work with users. This resource owner proposal is therefore only requesting access to the instruments.U.S. Department of Energy (DOE) user facilities excel at generating genomic (JGI) and proteomic (EMSL) data as well as solving protein structures (ALS, APS, EMSL NMR, LCLS, NSLS, SNS, SSRL). However, a rift exists in directly linking Office of Biological and Environmental Research (BER) systems biology and de novo structural biology approaches in a high-throughput manner using cryo-electron microscopy (cryo-EM), or for direct in situ label-free imaging of intact cells. A new cryogenic Transmission Electron Microscope (cryo-TEM) at EMSL could fill this gap specifically for BER users. To expand our onsite (cryo-EM) capability and accelerate science for the BER community, we propose to replace our existing 23-year-old JEOL 3000SFF cryo-TEM with a ThermoFisher Sciences Krios G3i for high-throughput screening and high-resolution cryo-EM data collection at EMSL to bridge the current capability divide and revitalize aging resources. The new instrument will be equipped with all modern best-option features such as grid autoloaders, hole-free phase plate, energy filter, bottom-mount CMOS detector, and Gatan K3 direct electron detection camera.
This state-of-the-art instrument will increase BER user impact and will help advance EMSL strategic science for achieving a holistic view of cellular ultrastructure needed to inform biodesign and understand biosystem dynamics, while simultaneously addressing the need for improved functional annotation called out in several recent DOE- BER reports. The refreshed cryo-EM capability at EMSL will permit BER researchers to access the instrument to directly benefit their research and to delve deep into the cell to understand the function of individual proteins identified as targets from proteomics. It will also help researchers to unveil the interactions of proteins occurring in the cell that are critical to function, but are currently invisible to classical biochemical, omics, and imaging approaches.
Project Details
Start Date
2020-10-02
End Date
2023-11-30
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members