1887

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Volume 101, Issue 11

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

Abstract

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.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): genotype 3 , hepatitis C virus , NS5A and sub-genomic replicon
Funding
This study was supported by the:
  • China Scholarship Council, http://dx.doi.org/10.13039/501100004543
  • FAPESP (Award 2018/04678-5)
  • FAPESP (Award 2016/03807-0)
  • Novo Nordisk Fonden, http://dx.doi.org/10.13039/501100009708 (Award NNF19OC0054518)
  • Medical Research Council, http://dx.doi.org/10.13039/501100000265 (Award MR/S001026/1)
© 2020 The Authors
  • 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.
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2020-09-08
2024-04-23
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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
J. Gen. Virol. 101, 1182 (2020); https://doi.org/10.1099/jgv.0.001486
/content/journal/jgv/10.1099/jgv.0.001486
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