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Inhibition of hepatitis C virus infection and expression in vitro and in vivo by recombinant adenovirus expressing short hairpin RNA
Author(s) -
Sakamoto Naoya,
Tanabe Yoko,
Yokota Takanori,
Satoh Kenichi,
SekineOsajima Yuko,
Nakagawa Mina,
Itsui Yasuhiro,
Tasaka Megumi,
Sakurai Yuki,
ChengHsin Chen,
Yano Masahiko,
Ohkoshi Shogo,
Aoyagi Yutaka,
Maekawa Shinya,
Enomoto Nobuyuki,
Kohara Michinori,
Watanabe Mamoru
Publication year - 2008
Publication title -
journal of gastroenterology and hepatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.214
H-Index - 130
eISSN - 1440-1746
pISSN - 0815-9319
DOI - 10.1111/j.1440-1746.2007.05076.x
Subject(s) - small hairpin rna , replicon , virology , transfection , retrovirus , microbiology and biotechnology , biology , genetic enhancement , hepatitis c virus , rna interference , small interfering rna , transduction (biophysics) , viral replication , virus , rna , cell culture , gene , plasmid , genetics , biochemistry
Background and Aim: We have reported previously that synthetic small interfering RNA (siRNA) and DNA‐based siRNA expression vectors efficiently and specifically suppress hepatitis C virus (HCV) replication in vitro . In this study, we investigated the effects of the siRNA targeting HCV‐RNA in vivo . Methods: We constructed recombinant retrovirus and adenovirus expressing short hairpin RNA (shRNA), and transfected into replicon‐expressing cells in vitro and transgenic mice in vivo . Results: Retroviral transduction of Huh7 cells to express shRNA and subsequent transfection of an HCV replicon into the cells showed that the cells had acquired resistance to HCV replication. Infection of cells expressing the HCV replicon with an adenovirus expressing shRNA resulted in efficient vector delivery and expression of shRNA, leading to suppression of the replicon in the cells by ∼10 −3 . Intravenous delivery of the adenovirus expressing shRNA into transgenic mice that can be induced to express HCV structural proteins by the Cre/ lox P switching system resulted in specific suppression of virus protein synthesis in the liver. Conclusion: Taken together, our results support the feasibility of utilizing gene targeting therapy based on siRNA and/or shRNA expression to counteract HCV replication, which might prove valuable in the treatment of hepatitis C.