Screening of hepatitis C virus inhibitors using genotype 1a HCV replicon cell lines

Curr Protoc Microbiol. 2011 Aug:Chapter 17:Unit17.7. doi: 10.1002/9780471729259.mc1707s22.

Abstract

Hepatitis C virus (HCV) replicons are the primary tools for identifying and evaluating anti-HCV compounds during drug discovery research. Genotype 1a and 1b are the most common genotypes in North America and Europe. These genotypes display significant genetic divergence (∼20% at the nucleotide level and ∼14% at the amino acid level), which can translate into marked differences in drug susceptibility. Thus, it is critical that potential therapeutics be assessed against both genotypes to ensure antiviral efficacy in the broad genotype 1 patient population. This unit describes assays for screening HCV inhibitors using replicons with a focus on genotype 1a. Specific protocols are provided for screening 1a replicons that encode the Renilla luciferase gene and for replicons that do not encode exogenous reporter genes, both in 96-well (manual) and in 384-well (high-throughput) formats.

MeSH terms

  • Antiviral Agents / pharmacology*
  • Cell Culture Techniques
  • Cell Line
  • Drug Evaluation, Preclinical / methods*
  • Genes, Reporter
  • Genotype
  • Hepacivirus / drug effects*
  • Hepacivirus / genetics*
  • Hepacivirus / metabolism
  • Humans
  • Microbial Sensitivity Tests / methods*
  • Replicon / drug effects*
  • Viral Nonstructural Proteins / antagonists & inhibitors
  • Viral Nonstructural Proteins / genetics
  • Viral Nonstructural Proteins / metabolism

Substances

  • Antiviral Agents
  • Viral Nonstructural Proteins