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Principal Investigator: ROBERT A DAVEY
Organization: WASHINGTON UNIVERSITY
Fiscal Year: 2023
Award: $371,762
Funding agency: National Institute of Allergy and Infectious Diseases
Core C – Project Summary/Abstract
Core C provides support for high biocontainment work to Research Projects 1 (RP01), RP02, RP03 and Core
B. Work is prioritized through Core A. Core personnel bring unique expertise in molecular biology, cell biology
and animal disease modeling for Ebola and Marburg viruses. The core maintains virus stocks, makes
recombinant viruses, provides cell lysates and RNA from infected cells and ensures they are free of infectious
material, evaluates effects of host factor loss or overexpression on infection and performs work with transgenic
rodents. These core activities provide needed protein material and an understanding of subcellular structures
made after virus infection for RP01, are required for evaluation of virus protein function and genome stem loop
structures in controlling transcription for RP02, and provide detailed analysis of infection kinetics and host-virus
co-association for RP03. To directly address impact of host factors on virus protein production, virus RNA
transcription and the substructures formed by each, we have developed a novel protein-antibody + vRNA-FISH
based staining technique that is quantitatively assessed by automated microscopy image analysis. This
approach is expected to provide important information for the needs of each project and Core B. Core C also
works closely with Core B in production and evaluation of novel antibodies by providing purified virus lysates
and performing microscopy evaluation of staining of infected cells. The project distinguishes itself in providing
high-level molecular biology ability at BSL4 that will be used to produce recombinant viruses predicted to have
altered replication kinetics. Furthermore, use of transgenic animals, provided through Core B, will be used to
evaluate roles in controlling disease. In so doing, Core C provides the necessary processes to evaluate host
factor roles from cells through to disease that enables identification of potential targets for therapy development.
Terms: <129 Mouse><Address><Allelism Test><Amino Acids><Animal Disease Models><Antibodies><BSL-4 facility><BSL4 facility><Behavior><Blood><Blood Reticuloendothelial System><Boston><Buffers><CRISPR><CRISPR/Cas system><Cell Body><Cells><Cellular biology><Clustered Regularly Interspaced Short Palindromic Repeats><Complementation Test><Cryo-electron Microscopy><Cryoelectron Microscopy><DNA Molecular Biology><Disease><Disease Outcome><Disease model><Disorder><Drugs><EBOV><Ebola virus><Ebola-like Viruses><Electron Cryomicroscopy><Emerging Communicable Diseases><Emerging Infectious Diseases><Ensure><Evaluation><Filoviridae><Filoviridae Infections><Filovirus><Frankfurt-Marburg Syndrome Virus><GRP94><Gene Transcription><Genetic Complementation Test><Genetic Transcription><Genome><Histologic><Histologically><Host Factor><Host Factor Protein><Human Resources><Image Analyses><Image Analysis><Immune Precipitation><Immunoblotting><Immunoprecipitation><Impact evaluation><Infection><Infectious Diseases / Laboratory><Infectious Diseases Research><Integration Host Factors><Intracellular Structure><KO mice><Kinetics><Knock-out><Knock-out Mice><Knockout><Knockout Mice><Liver><Manpower><Marburg><Marburg virus><Marburg-like Viruses><Marburgvirus><Measures><Medication><Methods><Mice><Mice Mammals><Microscopy><Modeling><Molecular><Molecular Biology><Mouse Strains><Murine><Mus><Non-Polyadenylated RNA><Nucleic Acids><Null Mouse><Outcome Measure><Patients><Pharmaceutic Preparations><Pharmaceutical Preparations><Phosphorylation><Process><Production><Protein Phosphorylation><Proteins><Publications><R-Series Research Projects><R01 Mechanism><R01 Program><RNA><RNA Expression><RNA Gene Products><Reagent><Recombinants><Research Grants><Research Project Grants><Research Projects><Research Resources><Resources><Ribonucleic Acid><Rodent><Rodentia><Rodents Mammals><Role><Science><Scientific Publication><Severities><Short interfering RNA><Single Crystal Diffraction><Small Interfering RNA><Source><Staining method><Stains><Structure><Subcellular structure><Sudan><Symptoms><System><Techniques><Testing><Training><Trans Test><Transcription><Transcription Regulation><Transcriptional Control><Transcriptional Regulation><Transgenic Animals><Transgenic Mice><Transgenic Organisms><Ubiquitilation><Ubiquitination><Ubiquitinoylation><Universities><Viral><Viral Antigens><Viral Diseases><Viral Gene Products><Viral Gene Proteins><Viral Proteins><Viral load measurement><Virion><Virus><Virus Diseases><Virus Particle><Western Blotting><Western Immunoblotting><Work><X Ray Crystallographies><X-Ray Crystallography><X-Ray Diffraction Crystallography><X-Ray/Neutron Crystallography><Xray Crystallography><aminoacid><biosafety level 4 facility><cell biology><complementation analysis><complementation approach><cryo-EM><cryoEM><cryogenic electron microscopy><depository><design><designing><develop therapy><disease control><disorder control><disorder model><drug/agent><ebolavirus><filovirus infections><forest><hepatic body system><hepatic organ system><image evaluation><image interpretation><in vivo><inhibitor><innovate><innovation><innovative><insight><intervention development><knock-down><knock-out animal><knockdown><knockout animal><knockout gene><measurable outcome><microscope imaging><microscopic imaging><microscopy imaging><mouse model><murine model><mutant><novel><novel virus><outcome measurement><overexpress><overexpression><pathogen><personnel><pharmacologic><programs><protein blotting><protein complex><protein expression><protein function><recombinant virus><repository><reverse genetics><siRNA><small molecule inhibitor><social role><stem><success><targeted drug therapy><targeted drug treatments><targeted therapeutic><targeted therapeutic agents><targeted therapy><targeted treatment><therapy development><transgenic><treatment development><trizol><ubiquination><ubiquitin conjugation><viral RNA><viral detection><viral infection><virus RNA><virus antigen><virus detection><virus infection><virus load><virus protein><virus-induced disease>