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Conditional Regulation of Gene Expression by Ligand-Induced Occlusion of a MicroRNA Target Sequence

RNA switches that modulate gene expression with small molecules have a number of scientific and clinical applications. Here, we describe a novel class of small regulatory on switches based on the ability of a ligand-bound aptamer to promote stem formation between a microRNA target sequence (miR-T) a...

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Autores principales: Mou, Huihui, Zhong, Guocai, Gardner, Matthew R., Wang, Haimin, Wang, Yi-Wen, Cheng, Dechun, Farzan, Michael
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society of Gene & Cell Therapy 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5993935/
https://www.ncbi.nlm.nih.gov/pubmed/29567311
http://dx.doi.org/10.1016/j.ymthe.2018.02.021
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author Mou, Huihui
Zhong, Guocai
Gardner, Matthew R.
Wang, Haimin
Wang, Yi-Wen
Cheng, Dechun
Farzan, Michael
author_facet Mou, Huihui
Zhong, Guocai
Gardner, Matthew R.
Wang, Haimin
Wang, Yi-Wen
Cheng, Dechun
Farzan, Michael
author_sort Mou, Huihui
collection PubMed
description RNA switches that modulate gene expression with small molecules have a number of scientific and clinical applications. Here, we describe a novel class of small regulatory on switches based on the ability of a ligand-bound aptamer to promote stem formation between a microRNA target sequence (miR-T) and a complementary competing strand. Two on switch architectures employing this basic concept were evaluated, differing in the location of a tetracycline aptamer and the region of a miR-21 target sequence (miR-21-T) masked by its competing strand. Further optimizations of miR-21-T and its competing strand resulted in tetracycline-regulated on switches that induced luciferase expression by 19-fold in HeLa cells. A similar switch design based on miR-122-T afforded 7-fold regulation when placed in tandem, indicating that this approach can be extended to additional miR-T. Optimized on switches introduced into adeno-associated virus (AAV) vectors afforded 10-fold regulation of two antiviral proteins in AAV-transduced cells. Our data demonstrate that small-molecule-induced occlusion of a miR-T can be used to conditionally regulate gene expression in mammalian cells and suggest that regulatory switches built on this principle can be used to dose expression of an AAV transgene.
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spelling pubmed-59939352019-05-02 Conditional Regulation of Gene Expression by Ligand-Induced Occlusion of a MicroRNA Target Sequence Mou, Huihui Zhong, Guocai Gardner, Matthew R. Wang, Haimin Wang, Yi-Wen Cheng, Dechun Farzan, Michael Mol Ther Original Article RNA switches that modulate gene expression with small molecules have a number of scientific and clinical applications. Here, we describe a novel class of small regulatory on switches based on the ability of a ligand-bound aptamer to promote stem formation between a microRNA target sequence (miR-T) and a complementary competing strand. Two on switch architectures employing this basic concept were evaluated, differing in the location of a tetracycline aptamer and the region of a miR-21 target sequence (miR-21-T) masked by its competing strand. Further optimizations of miR-21-T and its competing strand resulted in tetracycline-regulated on switches that induced luciferase expression by 19-fold in HeLa cells. A similar switch design based on miR-122-T afforded 7-fold regulation when placed in tandem, indicating that this approach can be extended to additional miR-T. Optimized on switches introduced into adeno-associated virus (AAV) vectors afforded 10-fold regulation of two antiviral proteins in AAV-transduced cells. Our data demonstrate that small-molecule-induced occlusion of a miR-T can be used to conditionally regulate gene expression in mammalian cells and suggest that regulatory switches built on this principle can be used to dose expression of an AAV transgene. American Society of Gene & Cell Therapy 2018-05-02 2018-02-27 /pmc/articles/PMC5993935/ /pubmed/29567311 http://dx.doi.org/10.1016/j.ymthe.2018.02.021 Text en © 2018 The Author(s) http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Original Article
Mou, Huihui
Zhong, Guocai
Gardner, Matthew R.
Wang, Haimin
Wang, Yi-Wen
Cheng, Dechun
Farzan, Michael
Conditional Regulation of Gene Expression by Ligand-Induced Occlusion of a MicroRNA Target Sequence
title Conditional Regulation of Gene Expression by Ligand-Induced Occlusion of a MicroRNA Target Sequence
title_full Conditional Regulation of Gene Expression by Ligand-Induced Occlusion of a MicroRNA Target Sequence
title_fullStr Conditional Regulation of Gene Expression by Ligand-Induced Occlusion of a MicroRNA Target Sequence
title_full_unstemmed Conditional Regulation of Gene Expression by Ligand-Induced Occlusion of a MicroRNA Target Sequence
title_short Conditional Regulation of Gene Expression by Ligand-Induced Occlusion of a MicroRNA Target Sequence
title_sort conditional regulation of gene expression by ligand-induced occlusion of a microrna target sequence
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5993935/
https://www.ncbi.nlm.nih.gov/pubmed/29567311
http://dx.doi.org/10.1016/j.ymthe.2018.02.021
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