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Title
  • Production of Vesicular Stomatitis Virus Glycoprotein-Pseudotyped Lentiviral Vector Is Enhanced by Ezrin Silencing
Creator

Izumida, Mai

Togawa, Kei

Hayashi, Hideki

Matsuyama, Toshifumi

Kubo, Yoshinao

Rights
    • © 2020 Izumida, Togawa, Hayashi, Matsuyama and Kubo. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted,provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
Subject
  • Other HIV-1
  • Other Gag protein
  • Other ezrin
  • Other phosphorylation
Description
Other
  • Human immunodeficiency virus type 1 (HIV-1)-based viral vector is widely used as a biomaterial to transfer a gene of interest into target cells in many biological study fields including gene therapy. Vesicular stomatitis virus glycoprotein (VSV-G)-containing HIV-1 vector much more efficiently transduces various mammalian cells than other viral envelope proteins-containing vectors. Understanding the mechanism would contribute to development of a novel method of efficient HIV-1 vector production. HIV-1 vector is generally constructed by transient transfection of human 293T or African green monkey COS7 cells. It was found in this study that HIV-1 Gag protein is constitutively digested in lysosomes of African green monkey cells. Surprisingly, VSV-G elevated HIV-1 Gag protein levels, suggesting that VSV-G protects Gag protein from the lysosomal degradation. Unphosphorylated ezrin, but not phosphorylated ezrin, was detected in COS7 cells, and ezrin silencing elevated Gag protein levels in the presence of VSV-G. Expression of unphosphorylated ezrin reduced Gag protein amounts. These results indicate that unphosphorylated ezrin proteins inhibit the VSV-G-mediated stabilization of HIV-1 Gag protein. Trafficking of HIV-1 Gag-associated intracellular vesicles may be controlled by ezrin. Finally, this study found that ezrin silencing yields higher amount of VSV-G-pseudotyped HIV-1 vector.
Other
  • identifier:Frontiers in Bioengineering and Biotechnology, 8, art.no.368; 2020
Other
  • identifier:22964185
PublisherFrontiers Media S.A.
Date Issued 2020-04-29
Languageeng
NIItypejournal article
VersiontypeVoR
Identifier URI http://hdl.handle.net/10069/40008
Relation
  • isIdenticalTo DOI https://doi.org/10.3389/fbioe.2020.00368
Journal
    • Frontiers in Bioengineering and Biotechnology
    8, 368
File
Oaidate2020-05-29T14:04:21Z