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Insights into the unique characteristics of hepatitis C virus genotype 3 revealed by development of a robust sub-genomic DBN3a replicon

Hepatitis C virus (HCV) is an important human pathogen causing 400 000 chronic liver disease-related deaths annually. Until recently, the majority of laboratory-based investigations into the biology of HCV have focused on the genotype 2 isolate, JFH-1, involving replicons and infectious cell culture...

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Autores principales: Ward, Joseph C., Bowyer, Sebastian, Chen, Shucheng, Fernandes Campos, Guilherme Rodrigues, Ramirez, Santseharay, Bukh, Jens, Harris, Mark
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Microbiology Society 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7879556/
https://www.ncbi.nlm.nih.gov/pubmed/32897181
http://dx.doi.org/10.1099/jgv.0.001486
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author Ward, Joseph C.
Bowyer, Sebastian
Chen, Shucheng
Fernandes Campos, Guilherme Rodrigues
Ramirez, Santseharay
Bukh, Jens
Harris, Mark
author_facet Ward, Joseph C.
Bowyer, Sebastian
Chen, Shucheng
Fernandes Campos, Guilherme Rodrigues
Ramirez, Santseharay
Bukh, Jens
Harris, Mark
author_sort Ward, Joseph C.
collection PubMed
description Hepatitis C virus (HCV) is an important human pathogen causing 400 000 chronic liver disease-related deaths annually. Until recently, the majority of laboratory-based investigations into the biology of HCV have focused on the genotype 2 isolate, JFH-1, involving replicons and infectious cell culture systems. However, genotype 2 is one of eight major genotypes of HCV and there is great sequence variation among these genotypes (>30 % nucleotide divergence). In this regard, genotype 3 is the second most common genotype and accounts for 30 % of global HCV cases. Further, genotype 3 is associated with both high levels of inherent resistance to direct-acting antiviral (DAA) therapy, and a more rapid progression to chronic liver diseases. Neither of these two attributes are fully understood, thus robust genotype 3 culture systems to unravel viral replication are required. Here we describe the generation of robust genotype 3 sub-genomic replicons (SGRs) based on the adapted HCV NS3-NS5B replicase from the DBN3a cell culture infectious clone. Such infectious cell culture-adaptive mutations could potentially promote the development of robust SGRs for other HCV strains and genotypes. The novel genotype 3 SGRs have been used both transiently and to establish stable SGR-harbouring cell lines. We show that these resources can be used to investigate aspects of genotype 3 biology, including NS5A function and DAA resistance. They will be useful tools for these studies, circumventing the need to work under the biosafety level 3 (BSL3) containment required in many countries.
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spelling pubmed-78795562021-02-12 Insights into the unique characteristics of hepatitis C virus genotype 3 revealed by development of a robust sub-genomic DBN3a replicon Ward, Joseph C. Bowyer, Sebastian Chen, Shucheng Fernandes Campos, Guilherme Rodrigues Ramirez, Santseharay Bukh, Jens Harris, Mark J Gen Virol Research Article Hepatitis C virus (HCV) is an important human pathogen causing 400 000 chronic liver disease-related deaths annually. Until recently, the majority of laboratory-based investigations into the biology of HCV have focused on the genotype 2 isolate, JFH-1, involving replicons and infectious cell culture systems. However, genotype 2 is one of eight major genotypes of HCV and there is great sequence variation among these genotypes (>30 % nucleotide divergence). In this regard, genotype 3 is the second most common genotype and accounts for 30 % of global HCV cases. Further, genotype 3 is associated with both high levels of inherent resistance to direct-acting antiviral (DAA) therapy, and a more rapid progression to chronic liver diseases. Neither of these two attributes are fully understood, thus robust genotype 3 culture systems to unravel viral replication are required. Here we describe the generation of robust genotype 3 sub-genomic replicons (SGRs) based on the adapted HCV NS3-NS5B replicase from the DBN3a cell culture infectious clone. Such infectious cell culture-adaptive mutations could potentially promote the development of robust SGRs for other HCV strains and genotypes. The novel genotype 3 SGRs have been used both transiently and to establish stable SGR-harbouring cell lines. We show that these resources can be used to investigate aspects of genotype 3 biology, including NS5A function and DAA resistance. They will be useful tools for these studies, circumventing the need to work under the biosafety level 3 (BSL3) containment required in many countries. Microbiology Society 2020-11 2020-09-08 /pmc/articles/PMC7879556/ /pubmed/32897181 http://dx.doi.org/10.1099/jgv.0.001486 Text en © 2020 The Authors http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License. This article was made open access via a Publish and Read agreement between the Microbiology Society and the corresponding author’s institution.
spellingShingle Research Article
Ward, Joseph C.
Bowyer, Sebastian
Chen, Shucheng
Fernandes Campos, Guilherme Rodrigues
Ramirez, Santseharay
Bukh, Jens
Harris, Mark
Insights into the unique characteristics of hepatitis C virus genotype 3 revealed by development of a robust sub-genomic DBN3a replicon
title Insights into the unique characteristics of hepatitis C virus genotype 3 revealed by development of a robust sub-genomic DBN3a replicon
title_full Insights into the unique characteristics of hepatitis C virus genotype 3 revealed by development of a robust sub-genomic DBN3a replicon
title_fullStr Insights into the unique characteristics of hepatitis C virus genotype 3 revealed by development of a robust sub-genomic DBN3a replicon
title_full_unstemmed Insights into the unique characteristics of hepatitis C virus genotype 3 revealed by development of a robust sub-genomic DBN3a replicon
title_short Insights into the unique characteristics of hepatitis C virus genotype 3 revealed by development of a robust sub-genomic DBN3a replicon
title_sort insights into the unique characteristics of hepatitis c virus genotype 3 revealed by development of a robust sub-genomic dbn3a replicon
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7879556/
https://www.ncbi.nlm.nih.gov/pubmed/32897181
http://dx.doi.org/10.1099/jgv.0.001486
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