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In vitro comparison of the internal ribosomal entry site activity from rodent hepacivirus and pegivirus and construction of pseudoparticles


Sims, Stuart; Michaelsen, Kevin; Burkhard, Sara; Fraefel, Cornel (2021). In vitro comparison of the internal ribosomal entry site activity from rodent hepacivirus and pegivirus and construction of pseudoparticles. Advances in Virology, 2021:5569844.

Abstract

The 5′ untranslated region (5′ UTR) of rodent hepacivirus (RHV) and pegivirus (RPgV) contains sequence homology to the HCV type III internal ribosome entry sites (IRES). Utilizing a monocistronic expression vector with an RNA polymerase I promoter to drive transcription, we show cell-specific IRES translation and regions within the IRES required for full functionality. Focusing on RHV, we further pseudotyped lentivirus with RHV and showed cell surface expression of the envelope proteins and transduction of murine hepatocytes and we then constructed full-length RHV and RPgV replicons with reporter genes. Using the replicon system, we show that the RHV NS3-4A protease cleaves a mitochondrial antiviral signaling protein reporter. However, liver-derived cells did not readily support the complete viral life cycle.

Abstract

The 5′ untranslated region (5′ UTR) of rodent hepacivirus (RHV) and pegivirus (RPgV) contains sequence homology to the HCV type III internal ribosome entry sites (IRES). Utilizing a monocistronic expression vector with an RNA polymerase I promoter to drive transcription, we show cell-specific IRES translation and regions within the IRES required for full functionality. Focusing on RHV, we further pseudotyped lentivirus with RHV and showed cell surface expression of the envelope proteins and transduction of murine hepatocytes and we then constructed full-length RHV and RPgV replicons with reporter genes. Using the replicon system, we show that the RHV NS3-4A protease cleaves a mitochondrial antiviral signaling protein reporter. However, liver-derived cells did not readily support the complete viral life cycle.

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Additional indexing

Item Type:Journal Article, not_refereed, original work
Communities & Collections:05 Vetsuisse Faculty > Veterinärwissenschaftliches Institut > Institute of Virology
Dewey Decimal Classification:570 Life sciences; biology
Scopus Subject Areas:Life Sciences > Virology
Health Sciences > Infectious Diseases
Uncontrolled Keywords:Virology, Infectious Diseases
Language:English
Date:29 July 2021
Deposited On:10 Aug 2021 08:37
Last Modified:25 Jun 2024 01:42
Publisher:Hindawi Publishing Corporation
ISSN:1687-8639
OA Status:Gold
Free access at:Publisher DOI. An embargo period may apply.
Publisher DOI:https://doi.org/10.1155/2021/5569844
  • Content: Published Version
  • Language: English
  • Licence: Creative Commons: Attribution 4.0 International (CC BY 4.0)