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KMID : 0545120060160101634
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Journal of Microbiology and Biotechnology
2006 Volume.16 No. 10 p.1634 ~ p.1639
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Inhibition of the Replication of Hepatitis C Virus Replicon with Nuclease-Resistant RNA Aptamers
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Shin Kyung-Sook
Lim Jong-Hoon
Kim Jung-Hye
Myung Hee-Joon
Lee Seong-Wook
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Abstract
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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 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.
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KEYWORD
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nuclease-resistant RNA aptamer, SELEX, HCV, HCV replicon, NS5B RNA replicase
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