Establishment of a hepatitis C virus subgenomic replicon derived from human hepatocytes infected in vitro

Nobuyuki Kato, Kazuo Sugiyama, Katsuyuki Namba, Hiromichi Dansako, Takashi Nakamura, Marika Takami, Kazuhito Naka, Akito Nozaki, Kunitada Shimotohno

Research output: Contribution to journalArticle

75 Citations (Scopus)

Abstract

The hepatitis C virus (HCV) replicon system is a potent tool for understanding the mechanisms of HCV replication and proliferation, and for the development of treatments for patients with HCV. Recently, we established an HCV subgenomic replicon (50-1) using HCV genome RNA obtained from the cultured human T cell line MT-2C infected with HCV (isolate 1B-1) in vitro. In order to further obtain other HCV replicons without difficulty, we generated a replicon RNA library derived from human non-neoplastic hepatocytes infected with HCV (isolate 1B-2) in vitro. Upon transfection of the generated RNA library to "cured cells," from which the 50-1 subgenomic replicon was eliminated by prolonged treatment with interferon-α, we successfully established a new HCV subgenomic replicon, 1B-2R1. We characterized 1B-2R1 replicon in terms of efficiency of replication, HCV sequence, and sensitivity to interferons. The results revealed that the replication level of the 1B-2R1 replicon was comparable to that of the 50-1 replicon. We also found that the 1B-2R1 replicon possessed an HCV sequence distinct from those of other replicons established to date, and that the 1B-2R1 replicon was sensitive to interferon-α, interferon-β, and interferon-γ. Taken together, present results indicate that the replicon RNA library generated using an in vitro HCV infection system is useful for the establishment of an HCV subgenomic replicon.

Original languageEnglish
Pages (from-to)756-766
Number of pages11
JournalBiochemical and Biophysical Research Communications
Volume306
Issue number3
DOIs
Publication statusPublished - Jul 4 2003

Keywords

  • Hepatitis C virus
  • Huh-7
  • Interferon
  • PH5CH8
  • Replication
  • Replicon

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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