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

被引：10

作者：

Dansako, Hiromichi ^{[1
]}

Ikeda, Masanori ^{[1
]}

Abe, Ken-ichi ^{[1
]}

Mori, Kyoko ^{[1
]}

Takemoto, Kazunori ^{[1
]}

Ariumi, Yasuo ^{[1
]}

Kato, Nobuyuki ^{[1
]}

机构：

[1] Okayama Univ, Grad Sch Med Dent & Pharmaceut Sci, Dept Mol Biol, Okayama 7008558, Japan

来源：

VIRUS RESEARCH | 2008年 / 137卷 / 01期

基金：

日本学术振兴会;

关键词：

hepatitis C virus; genome-length HCV RNA; living cell-based assay; green fluorescent protein; OGF7 assay system; anti-HCV reagents;

D O I：

10.1016/j.virusres.2008.06.001

中图分类号：

Q93 [微生物学];

学科分类号：

071005 [微生物学]; 100705 [微生物与生化药学];

摘要：

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-alpha treatment in a dose-dependent manner, and it correlated well with the HCV RNA concentration. We demonstrated that the interferon-alpha 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. (C) 2008 Elsevier B.V. All rights reserved.

引用

页码：72 / 79

页数：8

共 32 条

[1]

Cell culture-adaptive NS3 mutations required for the robust replication of genome-length hepatitis C virus RNA [J].