Split RNA switch orchestrates pre- and post-translational control to enable cell type-specific gene expression

Abstract RNA switch is a synthetic RNA-based technology that controls gene expression in response to cellular RNAs and proteins, thus enabling cell type-specific gene regulation and holding promise for gene therapy, regenerative medicine, and cell therapy. However, individual RNA switches often lack...

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Main Authors: Itsuki Abe, Hirohisa Ohno, Megumi Mochizuki, Karin Hayashi, Hirohide Saito
Format: Article
Language:English
Published: Nature Portfolio 2025-07-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-025-60392-2
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author Itsuki Abe
Hirohisa Ohno
Megumi Mochizuki
Karin Hayashi
Hirohide Saito
author_facet Itsuki Abe
Hirohisa Ohno
Megumi Mochizuki
Karin Hayashi
Hirohide Saito
author_sort Itsuki Abe
collection DOAJ
description Abstract RNA switch is a synthetic RNA-based technology that controls gene expression in response to cellular RNAs and proteins, thus enabling cell type-specific gene regulation and holding promise for gene therapy, regenerative medicine, and cell therapy. However, individual RNA switches often lack the specificity required for practical applications due to low ON/OFF ratios and difficulty in finding distinct and single biomolecule targets. To address these issues, we present “split RNA switches” that integrate outputs from multiple RNA switches by exploiting protein splicing. We show that split RNA switches significantly improve the ON/OFF ratio of microRNA-responsive ON switch system by canceling leaky OFF level in human cells. Using this approach, we achieve efficient cell purification using drug-resistance genes based on endogenous microRNA profiles and CRISPR-mediated genome editing with minimal off-target-cell effects. Additionally, we demonstrate RNA-based synthetic circuits using split RNA switches to enable the detection of multiple microRNAs and proteins with logical operations. Split RNA switches highlight the potential of post-translational processing as a versatile and comprehensive strategy for advancing mRNA-based therapeutic technologies.
format Article
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institution Kabale University
issn 2041-1723
language English
publishDate 2025-07-01
publisher Nature Portfolio
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series Nature Communications
spelling doaj-art-2b2533afd0b144c586d1f077d3fc25402025-08-20T04:01:35ZengNature PortfolioNature Communications2041-17232025-07-0116111810.1038/s41467-025-60392-2Split RNA switch orchestrates pre- and post-translational control to enable cell type-specific gene expressionItsuki Abe0Hirohisa Ohno1Megumi Mochizuki2Karin Hayashi3Hirohide Saito4Center for iPS Cell Research and Application, Kyoto University, 53 Kawahara-cho, Shogoin, Sakyo-kuCenter for iPS Cell Research and Application, Kyoto University, 53 Kawahara-cho, Shogoin, Sakyo-kuCenter for iPS Cell Research and Application, Kyoto University, 53 Kawahara-cho, Shogoin, Sakyo-kuCenter for iPS Cell Research and Application, Kyoto University, 53 Kawahara-cho, Shogoin, Sakyo-kuCenter for iPS Cell Research and Application, Kyoto University, 53 Kawahara-cho, Shogoin, Sakyo-kuAbstract RNA switch is a synthetic RNA-based technology that controls gene expression in response to cellular RNAs and proteins, thus enabling cell type-specific gene regulation and holding promise for gene therapy, regenerative medicine, and cell therapy. However, individual RNA switches often lack the specificity required for practical applications due to low ON/OFF ratios and difficulty in finding distinct and single biomolecule targets. To address these issues, we present “split RNA switches” that integrate outputs from multiple RNA switches by exploiting protein splicing. We show that split RNA switches significantly improve the ON/OFF ratio of microRNA-responsive ON switch system by canceling leaky OFF level in human cells. Using this approach, we achieve efficient cell purification using drug-resistance genes based on endogenous microRNA profiles and CRISPR-mediated genome editing with minimal off-target-cell effects. Additionally, we demonstrate RNA-based synthetic circuits using split RNA switches to enable the detection of multiple microRNAs and proteins with logical operations. Split RNA switches highlight the potential of post-translational processing as a versatile and comprehensive strategy for advancing mRNA-based therapeutic technologies.https://doi.org/10.1038/s41467-025-60392-2
spellingShingle Itsuki Abe
Hirohisa Ohno
Megumi Mochizuki
Karin Hayashi
Hirohide Saito
Split RNA switch orchestrates pre- and post-translational control to enable cell type-specific gene expression
Nature Communications
title Split RNA switch orchestrates pre- and post-translational control to enable cell type-specific gene expression
title_full Split RNA switch orchestrates pre- and post-translational control to enable cell type-specific gene expression
title_fullStr Split RNA switch orchestrates pre- and post-translational control to enable cell type-specific gene expression
title_full_unstemmed Split RNA switch orchestrates pre- and post-translational control to enable cell type-specific gene expression
title_short Split RNA switch orchestrates pre- and post-translational control to enable cell type-specific gene expression
title_sort split rna switch orchestrates pre and post translational control to enable cell type specific gene expression
url https://doi.org/10.1038/s41467-025-60392-2
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AT megumimochizuki splitrnaswitchorchestratespreandposttranslationalcontroltoenablecelltypespecificgeneexpression
AT karinhayashi splitrnaswitchorchestratespreandposttranslationalcontroltoenablecelltypespecificgeneexpression
AT hirohidesaito splitrnaswitchorchestratespreandposttranslationalcontroltoenablecelltypespecificgeneexpression