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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...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Microbiology Society
2020
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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. |
format | Online Article Text |
id | pubmed-7879556 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Microbiology Society |
record_format | MEDLINE/PubMed |
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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