A new living cell-based assay system for monitoring genome-length hepatitis C virus RNA replication

Hiromichi Dansako, Masanori Ikeda, Ken ichi Abe, Kyoko Mori, Kazunori Takemoto, Yasuo Ariumi, Nobuyuki Kato

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

We previously developed a cell-based luciferase reporter assay system for monitoring genome-length hepatitis C virus (HCV) RNA replication (OR6 assay system). Here, we aimed to develop a new living cell-based reporter assay system using enhanced green fluorescent protein (EGFP). Genome-length HCV RNAs encoding EGFP were introduced into a subline of HuH-7 cells and G418 selection was performed. One cloned cell line, OGF7, was successfully selected from among the several G418-resistant cell lines obtained, and the robust expression of HCV RNA and proteins in OGF7 cells was confirmed. The fluorescent intensity of OGF7 cells was decreased by interferon-α treatment in a dose-dependent manner, and it correlated well with the HCV RNA concentration. We demonstrated that the interferon-α sensitivity in the OGF7 assay system measuring the fluorescent intensity was equivalent to that of the OR6 assay system, and that the OGF7 assay system was useful for quantitative evaluation of anti-HCV reagents. The OGF7 assay system is expected to be the most time-saving and inexpensive assay system for high-throughput screening of anti-HCV reagents.

Original languageEnglish
Pages (from-to)72-79
Number of pages8
JournalVirus research
Volume137
Issue number1
DOIs
Publication statusPublished - Oct 2008

Keywords

  • Anti-HCV reagents
  • Genome-length HCV RNA
  • Green fluorescent protein
  • Hepatitis C virus
  • Living cell-based assay
  • OGF7 assay system

ASJC Scopus subject areas

  • Cancer Research
  • Virology
  • Infectious Diseases

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