Increased inhibitory ability of conjugated RNA aptamers against the HCV IRES |
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Authors: | Kunio Kikuchi Takuya Umehara Fumiko Nishikawa Tsunemi Hasegawa |
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Institution: | a Age Dimension Research Center, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Ibaraki 305-8566, Japan b Faculty of Science, Yamagata University, Yamagata 990-8560, Japan |
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Abstract: | Hepatitis C virus (HCV) translation begins within the internal ribosome entry site (IRES). We have previously isolated two RNA aptamers, 2-02 and 3-07, which specifically bind to domain II and domain III-IV of the HCV IRES, respectively, and inhibit IRES-dependent translation. To improve the function of these aptamers, we constructed two conjugated molecules of 2-02 and 3-07. These bound to the target RNA more efficiently than the two parental aptamers. Furthermore, they inhibited IRES-dependent translation about 10 times as efficiently as the 3-07 aptamer. This result indicates that combining aptamers for different target recognition sites potentiates the inhibition activity by enhancing the domain-binding efficiency. |
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Keywords: | HCV hepatitis C virus IRES internal ribosome entry site ME microchip electrophoresis PAGE polyacrylamide gel electrophoresis RRL rabbit reticulocyte lysate UTR untranslated region |
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