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A method for the generation of pseudovirus particles bearing SARS coronavirus spike protein in high yields

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Title: A method for the generation of pseudovirus particles bearing SARS coronavirus spike protein in high yields
Authors: Fujioka, Yoichiro Browse this author →KAKEN DB
Kashiwagi, Sayaka Browse this author
Yoshida, Aiko Browse this author
Satoh, Aya O. Browse this author
Fujioka, Mari Browse this author
Amano, Maho Browse this author
Yamauchi, Yohei Browse this author
Ohba, Yusuke Browse this author →KAKEN DB
Keywords: severe acute respiratory syndrome coronavirus (SARS-CoV)
vesicular stomatitis virus (VSV)
spike protein
Issue Date: 28-Apr-2022
Publisher: 一般社団法人 日本細胞生物学会(Japan Society for Cell Biology)
Journal Title: Cell structure and function
Volume: 47
Issue: 1
Start Page: 43
End Page: 53
Publisher DOI: 10.1247/csf.21047
Abstract: The ongoing severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic has threatened human health and the global economy. Development of additional vaccines and therapeutics is urgently required, but such development with live virus must be conducted with biosafety level 3 confinement. Pseudotyped viruses have been widely adopted for studies of virus entry and pharmaceutical development to overcome this restriction. Here we describe a modified protocol to generate vesicular stomatitis virus (VSV) pseudotyped with SARS-CoV or SARSCoV-2 spike protein in high yield. We found that a large proportion of pseudovirions produced with the conventional transient expression system lacked coronavirus spike protein at their surface as a result of inhibition of parental VSV infection by overexpression of this protein. Establishment of stable cell lines with an optimal expression level of coronavirus spike protein allowed the efficient production of progeny pseudoviruses decorated with spike protein. This improved VSV pseudovirus production method should facilitate studies of coronavirus entry and development of antiviral agents.
Type: article
Appears in Collections:医学院・医学研究院 (Graduate School of Medicine / Faculty of Medicine) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

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