{"title":"Structure-inhibition analysis of RNA aptamers that bind to HCV IRES.","authors":"Kunio Kikuchi, Takuya Umehara, Kotaro Fukuda, Joonsung Hwang, Atsushi Kuno, Tsunemi Hasegawa, Satoshi Nishikawa","doi":"10.1093/nass/3.1.291","DOIUrl":null,"url":null,"abstract":"The translation of HCV starts at the internal ribosomal entry site (IRES) within the 5' untranslated region and IRES is well-conserved in HCV strains. We developed a novel selection strategy using biotinylated oligonucleotide probe and obtained RNA aptamers that bind HCV IRES domain II and domain III-IV, respectively. Selected aptamers specifically bound to target sequence via RNA-RNA interactions. These aptamers inhibited IRES-depend translation in vitro. Especially, 3-07 aptamer, which bound domain IIId, showed strong inhibition. Structure/function relationship of these aptamers was analyzed by mutagenesis, RNase mapping and binding kinetics.","PeriodicalId":86149,"journal":{"name":"Nucleic acids research. Supplement (2001)","volume":" 3","pages":"291-2"},"PeriodicalIF":0.0000,"publicationDate":"2003-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1093/nass/3.1.291","citationCount":"13","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Nucleic acids research. Supplement (2001)","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1093/nass/3.1.291","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 13
Abstract
The translation of HCV starts at the internal ribosomal entry site (IRES) within the 5' untranslated region and IRES is well-conserved in HCV strains. We developed a novel selection strategy using biotinylated oligonucleotide probe and obtained RNA aptamers that bind HCV IRES domain II and domain III-IV, respectively. Selected aptamers specifically bound to target sequence via RNA-RNA interactions. These aptamers inhibited IRES-depend translation in vitro. Especially, 3-07 aptamer, which bound domain IIId, showed strong inhibition. Structure/function relationship of these aptamers was analyzed by mutagenesis, RNase mapping and binding kinetics.
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