title: Evolution of an inhibitory RNA aptamer against T7 RNA polymerase.
abstract: Aptamers are promising gene components that can be used for the construction of synthetic gene circuits. In this study, we isolated an RNA aptamer that specifically inhibits transcription of T7 RNA polymerase (RNAP). The 38-nucleotide aptamer, which was a shortened variant of an initial SELEX isolate, showed moderate inhibitory activity. By stepwise doped-SELEX, we isolated evolved variants with strong inhibitory activity. A 29-nucleotide variant of a doped-SELEX isolate showed 50% inhibitory concentration at 11 nM under typical in vitro transcription conditions. Pull-down experiments revealed that the aptamer inhibited the association of T7 RNAP with T7 promoter DNA.
authors: Department of Basic Medical Sciences, Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639, Japan.
authors: Shoji
authors: S
authors: Ohuchi
authors: Yusuke
authors: Y
authors: Mori
authors: Yoshikazu
authors: Y
authors: Nakamura
keywords: RNA aptamer; RNAP, RNA polymerase; SELEX; SELEX, systematic evolution of ligands by exponential enrichment; T7 RNA polymerase
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