Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Monitoring Viral Entry in Real-Time Using a Luciferase Recombinant Vesicular Stomatitis Virus Producing SARS-CoV-2, EBOV, LASV, CHIKV, and VSV Glycoproteins

Version 1 : Received: 24 November 2020 / Approved: 25 November 2020 / Online: 25 November 2020 (13:05:59 CET)

A peer-reviewed article of this Preprint also exists.

Lay Mendoza, M.F.; Acciani, M.D.; Levit, C.N.; Santa Maria, C.; Brindley, M.A. Monitoring Viral Entry in Real-Time Using a Luciferase Recombinant Vesicular Stomatitis Virus Producing SARS-CoV-2, EBOV, LASV, CHIKV, and VSV Glycoproteins. Viruses 2020, 12, 1457. Lay Mendoza, M.F.; Acciani, M.D.; Levit, C.N.; Santa Maria, C.; Brindley, M.A. Monitoring Viral Entry in Real-Time Using a Luciferase Recombinant Vesicular Stomatitis Virus Producing SARS-CoV-2, EBOV, LASV, CHIKV, and VSV Glycoproteins. Viruses 2020, 12, 1457.

Journal reference: Viruses 2020, 12, 1457
DOI: 10.3390/v12121457

Abstract

Viral entry is the first stage in the virus replication cycle and, for enveloped viruses, is mediated by virally encoded glycoproteins. Viral glycoproteins have different receptor affinities and triggering mechanisms. We employed vesicular stomatitis virus (VSV), a BSL-2 enveloped virus that can incorporate non-native glycoproteins, to examine the entry efficiencies of diverse viral glycoproteins. To compare glycoprotein-mediated entry efficiencies of: VSV G, SARS-CoV-2 S, EBOV GP, LASV GP, and CHIKV E we produced recombinant VSV (rVSV) viruses that produce the five glycoproteins. The rVSV virions encoded a nano luciferase-PEST (NLucP) reporter gene, which we used in combination with the live-cell substrate Endurazine™ to monitor viral entry kinetics in real time. Our data indicate that rVSV particles with glycoproteins that require more post-internalization priming typically demonstrate delayed entry in comparison to VSV G. In addition to determining the time required for each virus to complete entry, we also used our system to evaluate viral cell surface receptor preferences, monitor fusion, and elucidate endocytosis mechanisms. This system can be rapidly employed to examine diverse viral glycoproteins and their entry requirements.

Subject Areas

entry; kinetics; luciferase; real-time; live assay, vesicular stomatitis virus; Ebola; Lassa; chikungunya; coronavirus.

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