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Inhibition of the Replication of Hepatitis C Virus Replicon with Nuclease-Resistant RNA Aptamers  

Shin, Kyung-Sook (Department of Molecular Biology, Institute of Nanosensor and Biotechnology, Dankook University)
Lim, Jong-Hoon (Department of Molecular Biology, Institute of Nanosensor and Biotechnology, Dankook University)
Kim, Jung-Hye (Department of Molecular Biology, Institute of Nanosensor and Biotechnology, Dankook University)
Myung, Hee-Joon (Department of Bioscience and Biotechnology, Hankook University of Foreign Studies)
Lee, Seong-Wook (Department of Molecular Biology, Institute of Nanosensor and Biotechnology, Dankook University)
Publication Information
Journal of Microbiology and Biotechnology / v.16, no.10, 2006 , pp. 1634-1639 More about this Journal
Abstract
Hepatitis C virus (HCV)-encoded nonstructural protein 5B (NS5B) possesses RNA-dependent RNA polymerase activity, which is considered essential for viral proliferation. Thus, HCV NS5B is a good therapeutic target protein for the development of anti-HCV agents. In this study, we isolated two different kinds of nuclease-resistant RNA aptamers with 2'-fluoro pyrimidines against the HCV NS5B from a combinatorial RNA library with 40 nucleotide random sequences, using SELEX technology. The isolated RNA aptamers were observed to specifically and avidly bind the HCV NS5B with an apparent $K_d$ of 5 nM and 18 nM, respectively, in contrast with the original RNA library that hardly bound the target protein. Moreover, these aptamers could partially inhibit RNA synthesis of the HCV subgenomic replicon when transfected into Huh-7 hepatoma cell lines. These results suggest that the RNA aptamers selected in vitro could be useful not only as therapeutic agents of HCV infection but also as a powerful tool for the study of the HCV RNA-dependent RNA polymerase mechanism.
Keywords
HCV; HCV replicon; NS5B RNA replicase; nuclease-resistant RNA aptamer; SELEX;
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Times Cited By Web Of Science : 4  (Related Records In Web of Science)
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