Development of robust hepatitis C virus genotype 4 subgenomic replicons

Gastroenterology. 2013 Jan;144(1):59-61.e6. doi: 10.1053/j.gastro.2012.09.033. Epub 2012 Sep 20.

Abstract

Despite the prevalence of hepatitis C virus genotype 4, no replicon system is available for study of the genotype. To facilitate discovery and development of reagents against this virus, we synthesized and transcribed a genotype 4a subgenomic replicon and transfected Huh7-Lunet cells with it, which yielded very few colonies. However, when we used a new Huh-7-derived cell line, colony formation increased ∼70-fold. We identified multiple adaptive mutations in the virus's nonstructural 3 or 4A proteins that allowed the cells to maintain stable, genotype 4a luciferase-encoding replicons. Several classes of hepatitis C virus inhibitors had different antiviral effects on genotypes 4a vs 1b. The genotype 4a replicon system we created will aid in the development of treatment regimens for all genotypes of hepatitis C virus.

MeSH terms

  • Adaptation, Physiological / genetics
  • Antiviral Agents / pharmacology
  • Carrier Proteins / genetics
  • Cell Line, Tumor
  • Colony-Forming Units Assay
  • Genotype
  • Hepacivirus / genetics*
  • Humans
  • Intracellular Signaling Peptides and Proteins
  • Mutation
  • Plasmids
  • RNA Helicases / genetics
  • Replicon / drug effects
  • Replicon / genetics*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Viral Nonstructural Proteins / genetics

Substances

  • Antiviral Agents
  • Carrier Proteins
  • Intracellular Signaling Peptides and Proteins
  • NS3 protein, hepatitis C virus
  • NS4A cofactor peptide, Hepatitis C virus
  • Viral Nonstructural Proteins
  • NS-5 protein, hepatitis C virus
  • RNA Helicases