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Improved microRNA suppression by WPRE-linked tough decoy microRNA sponges

Our genes are post-transcriptionally regulated by microRNAs (miRNAs) inducing translational suppression and degradation of targeted mRNAs. Strategies to inhibit miRNAs in a spatiotemporal manner in a desired cell type or tissue, or at a desired developmental stage, can be crucial for understanding m...

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Autores principales: Hollensen, Anne Kruse, Thomsen, Rune, Bak, Rasmus O., Petersen, Charlotte Christie, Ermegaard, Eva R., Aagaard, Lars, Damgaard, Christian Kroun, Mikkelsen, Jacob Giehm
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory Press 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5513069/
https://www.ncbi.nlm.nih.gov/pubmed/28487381
http://dx.doi.org/10.1261/rna.061192.117
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author Hollensen, Anne Kruse
Thomsen, Rune
Bak, Rasmus O.
Petersen, Charlotte Christie
Ermegaard, Eva R.
Aagaard, Lars
Damgaard, Christian Kroun
Mikkelsen, Jacob Giehm
author_facet Hollensen, Anne Kruse
Thomsen, Rune
Bak, Rasmus O.
Petersen, Charlotte Christie
Ermegaard, Eva R.
Aagaard, Lars
Damgaard, Christian Kroun
Mikkelsen, Jacob Giehm
author_sort Hollensen, Anne Kruse
collection PubMed
description Our genes are post-transcriptionally regulated by microRNAs (miRNAs) inducing translational suppression and degradation of targeted mRNAs. Strategies to inhibit miRNAs in a spatiotemporal manner in a desired cell type or tissue, or at a desired developmental stage, can be crucial for understanding miRNA function and for pushing forward miRNA suppression as a feasible rationale for genetic treatment of disease. For such purposes, RNA polymerase II (RNA Pol II)-transcribed tough decoy (TuD) miRNA inhibitors are particularly attractive. Here, we demonstrate augmented miRNA suppression capacity of TuD RNA hairpins linked to the Woodchuck hepatitis virus post-transcriptional regulatory element (WPRE). This effect is position-dependent and evident only when the WPRE is positioned upstream of the TuD. In accordance, inclusion of the WPRE does not change nuclear export, translation, total levels of TuD-containing RNA transcripts, or cytoplasmic P-body localization, suggesting that previously reported WPRE functions are negligible for improved TuD function. Notably, deletion analysis of TuD-fused WPRE unveils truncated WPRE variants resulting in optimized miRNA suppression. Together, our findings add to the guidelines for production of WPRE-supported anti-miRNA TuDs.
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spelling pubmed-55130692018-08-01 Improved microRNA suppression by WPRE-linked tough decoy microRNA sponges Hollensen, Anne Kruse Thomsen, Rune Bak, Rasmus O. Petersen, Charlotte Christie Ermegaard, Eva R. Aagaard, Lars Damgaard, Christian Kroun Mikkelsen, Jacob Giehm RNA Article Our genes are post-transcriptionally regulated by microRNAs (miRNAs) inducing translational suppression and degradation of targeted mRNAs. Strategies to inhibit miRNAs in a spatiotemporal manner in a desired cell type or tissue, or at a desired developmental stage, can be crucial for understanding miRNA function and for pushing forward miRNA suppression as a feasible rationale for genetic treatment of disease. For such purposes, RNA polymerase II (RNA Pol II)-transcribed tough decoy (TuD) miRNA inhibitors are particularly attractive. Here, we demonstrate augmented miRNA suppression capacity of TuD RNA hairpins linked to the Woodchuck hepatitis virus post-transcriptional regulatory element (WPRE). This effect is position-dependent and evident only when the WPRE is positioned upstream of the TuD. In accordance, inclusion of the WPRE does not change nuclear export, translation, total levels of TuD-containing RNA transcripts, or cytoplasmic P-body localization, suggesting that previously reported WPRE functions are negligible for improved TuD function. Notably, deletion analysis of TuD-fused WPRE unveils truncated WPRE variants resulting in optimized miRNA suppression. Together, our findings add to the guidelines for production of WPRE-supported anti-miRNA TuDs. Cold Spring Harbor Laboratory Press 2017-08 /pmc/articles/PMC5513069/ /pubmed/28487381 http://dx.doi.org/10.1261/rna.061192.117 Text en © 2017 Hollensen et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society http://creativecommons.org/licenses/by-nc/4.0/ This article is distributed exclusively by the RNA Society for the first 12 months after the full-issue publication date (see http://rnajournal.cshlp.org/site/misc/terms.xhtml). After 12 months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.
spellingShingle Article
Hollensen, Anne Kruse
Thomsen, Rune
Bak, Rasmus O.
Petersen, Charlotte Christie
Ermegaard, Eva R.
Aagaard, Lars
Damgaard, Christian Kroun
Mikkelsen, Jacob Giehm
Improved microRNA suppression by WPRE-linked tough decoy microRNA sponges
title Improved microRNA suppression by WPRE-linked tough decoy microRNA sponges
title_full Improved microRNA suppression by WPRE-linked tough decoy microRNA sponges
title_fullStr Improved microRNA suppression by WPRE-linked tough decoy microRNA sponges
title_full_unstemmed Improved microRNA suppression by WPRE-linked tough decoy microRNA sponges
title_short Improved microRNA suppression by WPRE-linked tough decoy microRNA sponges
title_sort improved microrna suppression by wpre-linked tough decoy microrna sponges
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5513069/
https://www.ncbi.nlm.nih.gov/pubmed/28487381
http://dx.doi.org/10.1261/rna.061192.117
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