title: Development of robust hepatitis C virus genotype 4 subgenomic replicons.
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.
authors.affiliation: Biology Department, Gilead Sciences, Foster City, California, USA.
authors.forename: Betty
authors.initials: B
authors.lastname: Peng
authors.forename: Mei
authors.initials: M
authors.lastname: Yu
authors.forename: Simin
authors.initials: S
authors.lastname: Xu
authors.forename: Yu-Jen
authors.initials: YJ
authors.lastname: Lee
authors.forename: Yang
authors.initials: Y
authors.lastname: Tian
authors.forename: Huiling
authors.initials: H
authors.lastname: Yang
authors.forename: Katie
authors.initials: K
authors.lastname: Chan
authors.forename: Hongmei
authors.initials: H
authors.lastname: Mo
authors.forename: John
authors.initials: J
authors.lastname: McHutchison
authors.forename: William
authors.initials: W
authors.lastname: Delaney
authors.forename: Guofeng
authors.initials: G
authors.lastname: Cheng
mesh_terms: D000222:Adaptation, Physiological / Q000235:genetics; D000998:Antiviral Agents / Q000494:pharmacology; D002352:Carrier Proteins / Q000235:genetics; D045744:Cell Line, Tumor; D003114:Colony-Forming Units Assay; D005838:Genotype; D016174:Hepacivirus / Q000235:genetics*; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D009154:Mutation; D010957:Plasmids; D020365:RNA Helicases / Q000235:genetics; D012093:Replicon / Q000187:drug effects / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017361:Viral Nonstructural Proteins / Q000235:genetics; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases; D012324:RNA-Dependent RNA Polymerase
chemical_list: D000998:Antiviral Agents; D002352:Carrier Proteins; D047908:Intracellular Signaling Peptides and Proteins; D020365:RNA Helicases; D017361:Viral Nonstructural Proteins; C084422:NS3 protein, hepatitis C virus; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases; C455041:NS4A cofactor peptide, Hepatitis C virus; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase