Evolution of an inhibitory RNA aptamer against T7 RNA polymerase

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 isolat...

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Detalles Bibliográficos
Autores principales: Ohuchi, Shoji, Mori, Yusuke, Nakamura, Yoshikazu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2012
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3642155/
https://www.ncbi.nlm.nih.gov/pubmed/23650601
http://dx.doi.org/10.1016/j.fob.2012.07.004
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author Ohuchi, Shoji
Mori, Yusuke
Nakamura, Yoshikazu
author_facet Ohuchi, Shoji
Mori, Yusuke
Nakamura, Yoshikazu
author_sort Ohuchi, Shoji
collection PubMed
description 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.
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spelling pubmed-36421552013-05-06 Evolution of an inhibitory RNA aptamer against T7 RNA polymerase Ohuchi, Shoji Mori, Yusuke Nakamura, Yoshikazu FEBS Open Bio Article 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. Elsevier 2012-07-20 /pmc/articles/PMC3642155/ /pubmed/23650601 http://dx.doi.org/10.1016/j.fob.2012.07.004 Text en © 2012 Published by Elsevier B.V. on behalf of Federation of European Biochemical Societies. This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-No Derivative Works License, which permits non- commercial use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Article
Ohuchi, Shoji
Mori, Yusuke
Nakamura, Yoshikazu
Evolution of an inhibitory RNA aptamer against T7 RNA polymerase
title Evolution of an inhibitory RNA aptamer against T7 RNA polymerase
title_full Evolution of an inhibitory RNA aptamer against T7 RNA polymerase
title_fullStr Evolution of an inhibitory RNA aptamer against T7 RNA polymerase
title_full_unstemmed Evolution of an inhibitory RNA aptamer against T7 RNA polymerase
title_short Evolution of an inhibitory RNA aptamer against T7 RNA polymerase
title_sort evolution of an inhibitory rna aptamer against t7 rna polymerase
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3642155/
https://www.ncbi.nlm.nih.gov/pubmed/23650601
http://dx.doi.org/10.1016/j.fob.2012.07.004
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AT moriyusuke evolutionofaninhibitoryrnaaptameragainstt7rnapolymerase
AT nakamurayoshikazu evolutionofaninhibitoryrnaaptameragainstt7rnapolymerase

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