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A novel luciferase and GFP dual reporter virus for rapid and convenient evaluation of hepatitis C virus replication

Wu, Yang, Liao, Qingjiao, Yang, Rongge, Chen, Xinwen, Chen, Xulin
Virus research 2011 v.155 no.2 pp. 406-414
Hepatitis C virus, RNA, antiviral agents, cost effectiveness, drugs, fluorescence, green fluorescent protein, luciferase, monitoring, open reading frames, reporter genes, screening, viral load, viral nonstructural proteins, virion, virus replication, viruses
Herein, we describe the development of a monocistronic dual reporter virus for monitoring hepatitis C virus (HCV) replication. The recombinant construct encodes for the humanized Renilla luciferase (hRLuc) reporter gene inserted upstream of the viral open reading frame and a green fluorescent protein (GFP) gene inserted into the C-terminus of non-structural protein 5A (NS5A) of the JFH1 viral genome. The viral RNA replicated efficiently in transfected cells and infectious virions could be produced without obvious attenuation of viral replication. The viral titer of the dual reporter virus was comparable to that of single reporter viruses. The expression levels of these two reporter genes correlated well with HCV replication in the presence or absence of antiviral agents. Moreover, because of the direct visibility of GFP fluorescence and the correlation between GFP positive cell numbers and hRLuc activity, the optimal time for measuring hRLuc activity was determined. This novel infectious system is a time saving and cost effective method for studying the interaction between viruses and host cells as well as for screening anti-HCV drugs.