Artificial turn-on riboswitch to control target gene expression using a wild-type riboswitch splicing mechanism.

Takahiro Yamauchi, Takafumi Kubodera, Daisuke Miyoshi, Naoki Sugimoto, Shuji Hirohata

Journal of bioscience and bioengineering 2021 Feb

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The thiamine pyrophospate (TPP)-dependent thiA riboswitch in Aspergillus oryzae regulates alternative mRNA splicing via TPP-riboswitch binding to reduce protein production. Based on the sequences involved in alternative splicing found in Neurospora crassa, we identified unique sequences that are thought to play a role in the alternative splicing of the thiA riboswitch. Based on the mechanism of alternative splicing regulated by the thiA riboswitch, we constructed a new TPP-dependent artificial riboswitch, which unlike the wild-type riboswitch, promotes, rather than inhibits, gene expression. We found that a target gene controlled by this turn-on artificial riboswitch can be sufficiently expressed for practical use in A. oryzae. The artificial riboswitch upregulates the target genes via TPP and can be applied as a practical tool for gene regulation. Copyright © 2020 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Citation

Takahiro Yamauchi, Takafumi Kubodera, Daisuke Miyoshi, Naoki Sugimoto, Shuji Hirohata. Artificial turn-on riboswitch to control target gene expression using a wild-type riboswitch splicing mechanism. Journal of bioscience and bioengineering. 2021 Feb;131(2):115-123