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Site-specific biotinylation of RNA molecules by transcription using unnatural base pairs.

ABSTRACT: Direct site-specific biotinylation of RNA molecules was achieved by specific transcription mediated by unnatural base pairs. Unnatural base pairs between 2-amino-6-(2-thienyl)purine (denoted by s) and 2-oxo(1H)pyridine (denoted by y), or 2-amino-6-(2-thiazolyl)purine (denoted as v) and y specifically function in T7 transcription. Using these unnatural base pairs, the substrate of biotinylated-y (Bio-yTP) was selectively incorporated into RNA, opposite s or v in the DNA templates, by T7 RNA polymerase. This method was applied to the immobilization of an RNA aptamer on sensor chips, and the aptamer accurately recognized its target protein. This direct site-specific biotinylation will provide a tool for RNA-based biotechnologies.

SUBMITTER: Moriyama K

PROVIDER: S-EPMC1188086 | biostudies-literature | 2005 Aug

REPOSITORIES: biostudies-literature

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Site-specific biotinylation of RNA molecules by transcription using unnatural base pairs.

Moriyama Kei K Kimoto Michiko M Mitsui Tsuneo T Yokoyama Shigeyuki S Hirao Ichiro I

Nucleic acids research 20050819 15

Direct site-specific biotinylation of RNA molecules was achieved by specific transcription mediated by unnatural base pairs. Unnatural base pairs between 2-amino-6-(2-thienyl)purine (denoted by s) and 2-oxo(1H)pyridine (denoted by y), or 2-amino-6-(2-thiazolyl)purine (denoted as v) and y specifically function in T7 transcription. Using these unnatural base pairs, the substrate of biotinylated-y (Bio-yTP) was selectively incorporated into RNA, opposite s or v in the DNA templates, by T7 RNA polym ...[more]

PMID: 16113238

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