Cryo-Electron Microscopy Research Infrastructure
Lead Research Organisation:
University of Oxford
Department Name: UNLISTED
Abstract
The Oxford Centre for Particle Imaging at the Wellcome Trust Centre for Human Genetics is pivotal for pan-departmental access and training in biological electron cryo microscopy (cryoEM) and electron cryo tomography (cryoET) for the 300+ structural biology research community in Oxford. State-of-the-art cryoEM and cryoET can image the molecular mechanisms by which human cells, disease pathogens and vaccines work. Such studies can pinpoint how differences in the genetic makeup of individuals contribute to a wide range of medical conditions through effects at the cellular and molecular level. Likewise, the design of vaccines to combat emergent viruses can be guided by mapping the surface details of the virus that are best recognised by our immune system. This grant application is to upgrade instrumentation for state-of-the-art cryoEM and cryoET in OPIC. OPIC is
unique in Europe in having the capability, when necessary, to run cryoEM and cryoET data collection in containment. OPIC, through its association with the UK national facility, eBIC (electron Bio-Imaging Centre) at Diamond Light Source, provides access for such experiments to the broader national and international user community. On-going analyses on viruses and vaccines important to human health worldwide will benefit from the requested equipment.
unique in Europe in having the capability, when necessary, to run cryoEM and cryoET data collection in containment. OPIC, through its association with the UK national facility, eBIC (electron Bio-Imaging Centre) at Diamond Light Source, provides access for such experiments to the broader national and international user community. On-going analyses on viruses and vaccines important to human health worldwide will benefit from the requested equipment.
Technical Summary
The Oxford Centre for Particle Imaging (OPIC) at the Wellcome Trust Centre for Human Genetics (WTCHG) plays the leading role for biological cryo electron microscopy and tomography (cryoEM and cryoET) at Oxford University. OPIC also offers a unique capability of contained microscopy as part of eBIC, the national facility. The high-end cryoEM instrument in OPIC is a 12-year-old FEI F30
‘Polara’, which has become a bottleneck for our science due to its unreliability, low throughput and lack of ability to integrate latest technologies.
The requested Krios cryoEM will:
- Underpin the research of multiple groups within the WTCHG, that of the Structural Genomics Consortium (SGC) and other neighbouring biomedical researchers on the Old Road Campus.
- Serve as a lynch pin in the pan-departmental strategy for biological cryoEM across the University of Oxford.
- Provide access to high-end cryoEM and cryoET for eBIC users with projects requiring ACDP category 3 and DEFRA 4 levels of containment.
Research that will benefit includes:
- ‘structural vaccinology’ combining deep/single cell sequencing of the humoral response to emergent viruses with cryoEM mapping of epitopes.
- analyses of cell-cell signalling systems and virus-cell interactions using sub-tomogram averaging and whole cell cryoET combined with correlative light microscopy.
‘Polara’, which has become a bottleneck for our science due to its unreliability, low throughput and lack of ability to integrate latest technologies.
The requested Krios cryoEM will:
- Underpin the research of multiple groups within the WTCHG, that of the Structural Genomics Consortium (SGC) and other neighbouring biomedical researchers on the Old Road Campus.
- Serve as a lynch pin in the pan-departmental strategy for biological cryoEM across the University of Oxford.
- Provide access to high-end cryoEM and cryoET for eBIC users with projects requiring ACDP category 3 and DEFRA 4 levels of containment.
Research that will benefit includes:
- ‘structural vaccinology’ combining deep/single cell sequencing of the humoral response to emergent viruses with cryoEM mapping of epitopes.
- analyses of cell-cell signalling systems and virus-cell interactions using sub-tomogram averaging and whole cell cryoET combined with correlative light microscopy.
Organisations
Publications
Bahar MW
(2022)
A conserved glutathione binding site in poliovirus is a target for antivirals and vaccine stabilisation.
in Communications biology
Avanzato V
(2024)
A monoclonal antibody targeting the Nipah virus fusion glycoprotein apex imparts protection from disease
in Journal of Virology
Howlett MG
(2022)
An autonomously oscillating supramolecular self-replicator.
in Nature chemistry
McCorvie TJ
(2024)
Architecture and regulation of filamentous human cystathionine beta-synthase.
in Nature communications
Duyvesteyn HME
(2021)
Bacteriophage PRD1 as a nanoscaffold for drug loading.
in Nanoscale
Ni T
(2023)
ChAdOx1 COVID vaccines express RBD open prefusion SARS-CoV-2 spikes on the cell surface
in iScience
Shah P
(2023)
Characterization of the rotavirus assembly pathway in situ using cryoelectron tomography
in Cell Host & Microbe
Chi G
(2022)
Cryo-EM structure of the human Kv3.1 channel reveals gating control by the cytoplasmic T1 domain.
in Nature communications
Yu X
(2022)
Cryo-EM structures of perforin-2 in isolation and assembled on a membrane suggest a mechanism for pore formation.
in The EMBO journal
Kus K
(2025)
DSIF factor Spt5 coordinates transcription, maturation and exoribonucleolysis of RNA polymerase II transcripts
in Nature Communications
Renner M
(2021)
Flavivirus maturation leads to the formation of an occupied lipid pocket in the surface glycoproteins.
in Nature communications
Ni T
(2023)
Intrinsically disordered CsoS2 acts as a general molecular thread for a-carboxysome shell assembly.
in Nature communications
Bahar M
(2021)
Mammalian expression of virus-like particles as a proof of principle for next generation polio vaccines
in npj Vaccines
Bahar MW
(2021)
Mammalian expression of virus-like particles as a proof of principle for next generation polio vaccines.
in NPJ vaccines
Keown JR
(2022)
Mapping inhibitory sites on the RNA polymerase of the 1918 pandemic influenza virus using nanobodies.
in Nature communications
Uchanski T
(2021)
Megabodies expand the nanobody toolkit for protein structure determination by single-particle cryo-EM.
in Nature methods
Krebs A
(2023)
Molecular architecture and conservation of an immature human endogenous retrovirus
in Nature Communications
McCorvie TJ
(2022)
Molecular basis for the regulation of human glycogen synthase by phosphorylation and glucose-6-phosphate.
in Nature structural & molecular biology
Pražák V
(2024)
Molecular plasticity of herpesvirus nuclear egress analysed in situ.
in Nature microbiology
Waheed AA
(2023)
Neutral sphingomyelinase 2 is required for HIV-1 maturation.
in Proceedings of the National Academy of Sciences of the United States of America
Chi G
(2021)
Phospho-regulation, nucleotide binding and ion access control in potassium-chloride cotransporters.
in The EMBO journal
Chi G
(2021)
Phospho-regulation, nucleotide binding and ion access control in potassium-chloride cotransporters.
in The EMBO journal
Nutalai R
(2022)
Potent cross-reactive antibodies following Omicron breakthrough in vaccinees.
in Cell
Sherry L
(2022)
Production and Characterisation of Stabilised PV-3 Virus-like Particles Using Pichia pastoris.
in Viruses
Robinson RA
(2021)
Simultaneous binding of Guidance Cues NET1 and RGM blocks extracellular NEO1 signaling.
in Cell
Kelly JJ
(2022)
Snapshots of actin and tubulin folding inside the TRiC chaperonin.
in Nature structural & molecular biology
Keown J
(2024)
Structural and functional characterization of the interaction between the influenza A virus RNA polymerase and the CTD of host RNA polymerase II.
in Journal of virology
Mycroft-West CJ
(2024)
Structural and mechanistic characterization of bifunctional heparan sulfate N-deacetylase-N-sulfotransferase 1.
in Nature communications
Keown J
(2024)
Structural characterization of the full-length Hantaan virus polymerase
in PLOS Pathogens
Huang S
(2024)
Structural insights into the mechanisms of urea permeation and distinct inhibition modes of urea transporters
in Nature Communications
Li HZ
(2024)
Structure and function of the SIT1 proline transporter in complex with the COVID-19 receptor ACE2.
in Nature communications
Hou Z
(2023)
Structure of native chromatin fibres revealed by Cryo-ET in situ.
in Nature communications
Whitehead J
(2023)
Structure of the N-RNA/P interface indicates mode of L/P recruitment to the nucleocapsid of human metapneumovirus
in Nature Communications
Balikçi E
(2024)
Structure of the Nipah virus polymerase complex
in The EMBO Journal
Ng WM
(2022)
Structure of trimeric pre-fusion rabies virus glycoprotein in complex with two protective antibodies.
in Cell host & microbe
Coupland CE
(2021)
Structure, mechanism, and inhibition of Hedgehog acyltransferase.
in Molecular cell
Huang KA
(2022)
Structures and therapeutic potential of anti-RBD human monoclonal antibodies against SARS-CoV-2.
in Theranostics
Staller E
(2024)
Structures of H5N1 influenza polymerase with ANP32B reveal mechanisms of genome replication and host adaptation.
in Nature communications
Fan H
(2019)
Structures of influenza A virus RNA polymerase offer insight into viral genome replication.
in Nature
Zhou D
(2022)
Switching of Receptor Binding Poses between Closely Related Enteroviruses.
in Viruses
Liu C
(2022)
The antibody response to SARS-CoV-2 Beta underscores the antigenic distance to other variants
in Cell Host & Microbe
Dejnirattisai W
(2021)
The antigenic anatomy of SARS-CoV-2 receptor binding domain.
in Cell
Liang Q
(2024)
The binding and mechanism of a positive allosteric modulator of Kv3 channels
in Nature Communications
Kleinpeter A
(2023)
The Effect of Inositol Hexakisphosphate on HIV-1 Particle Production and Infectivity can be Modulated by Mutations that Affect the Stability of the Immature Gag Lattice
in Journal of Molecular Biology