Production of Filovirus Glycoprotein-Pseudotyped Vesicular Stomatitis Virus for Study of Filovirus Entry Mechanisms

Rachel B Brouillette; Wendy Maury

doi:10.1007/978-1-4939-7116-9_4

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Journal article

Production of Filovirus Glycoprotein-Pseudotyped Vesicular Stomatitis Virus for Study of Filovirus Entry Mechanisms

Rachel B Brouillette and Wendy Maury

Methods in molecular biology (Clifton, N.J.), Vol.1628, pp.53-63

2017

DOI: 10.1007/978-1-4939-7116-9_4

PMCID: PMC7787361

PMID: 28573610

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Abstract

Members of the family Filoviridae are filamentous, enveloped, and nonsegmented negative-stranded RNA viruses that can cause severe hemorrhagic disease in humans and nonhuman primates with high mortality rates. Current efforts to analyze the structure and biology of these viruses as well as the development of antivirals have been hindered by the necessity of biosafety level 4 containment (BSL4). Here, we outline how to produce and work with Ebola virus glycoprotein bearing vesicular stomatitis virus (VSV) pseudovirions. These pseudovirions can be safely used to evaluate early steps of the filovirus life cycle without need for BSL4 containment. Virus gene expression in the transduced cells is easy to assess since the pseudovirions encode a reporter gene in place of the VSV G glycoprotein gene. Adoption of VSV for use as a pseudovirion system for filovirus GP has significantly expanded access for researchers to study specific aspects of the viral life cycle outside of BSL4 containment and has allowed substantial growth of filovirus research.

Vesicular Stomatitis - pathology

Viral Envelope Proteins - genetics

RNA Viruses - pathogenicity

Humans

RNA Viruses - genetics

Virion - genetics

Ebolavirus - pathogenicity

Genes, Reporter - genetics

Containment of Biohazards

Ebolavirus - genetics

Virus Internalization

Membrane Glycoproteins - genetics

Vesiculovirus - pathogenicity

Vesiculovirus - genetics

Vesicular Stomatitis - virology

Details

Title: Subtitle: Production of Filovirus Glycoprotein-Pseudotyped Vesicular Stomatitis Virus for Study of Filovirus Entry Mechanisms
Creators: Rachel B Brouillette - Department of Microbiology, University of Iowa, Iowa City, IA, USA
Wendy Maury - Department of Microbiology, University of Iowa, Iowa City, IA, USA. wendy-maury@uiowa.edu
Resource Type: Journal article
Publication Details: Methods in molecular biology (Clifton, N.J.), Vol.1628, pp.53-63
DOI: 10.1007/978-1-4939-7116-9_4
PMID: 28573610
PMCID: PMC7787361
NLM abbreviation: Methods Mol Biol
eISBN: 9781493971169; 1493971166
ISSN: 1064-3745
eISSN: 1940-6029
Grant note: R21 AI123616 / NIAID NIH HHS T32 AI007533 / NIAID NIH HHS
Language: English
Date published: 2017
Academic Unit: Microbiology and Immunology
Record Identifier: 9984083884002771

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