Analysis of replication of Hepatitis C Virus using highly efficient replication system

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Hepatitis C virus (HCV) is a major public health problem, infecting an estimated 170 million people worldwide. Current therapy for HCV-related chronic hepatitis is based on the use of interferon. However, virus clearance rates are insufficient. Investigations to develop the anti-viral therapy or to understand the life cycle of this virus have been hampered by the lack of viral culture systems. We isolated the JFH-1 strain from a patient with fulminant hepatitis, and the JFH-1 subgenomic replicon could replicate efficiently in culture cell without adaptive mutation. Using this JFH-1 strain, we developed highly efficient HCV replication system in culture cells. The full-length JFH-1 RNA was transfected into Huh7 cells. Subsequently, viral RNA efficiently replicated in transfected cells and viral particles were secreted. Secreted viral particle exhibited spherical shape by electron-microscopic analysis and was separated in the fraction eith 1.15g/ml density. Furthermore, secreted virus displayed infectivity for naive Huh7 cells. In addition, we have also developed an replicon system using full-genomic JFH-1 genome. Both systems provide powerful tools for studying the viral life cycle and constructing anti-viral strategies.