Cargando…
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...
Autores principales: | , , , , , , , |
---|---|
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 |
_version_ | 1783250589230039040 |
---|---|
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. |
format | Online Article Text |
id | pubmed-5513069 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Cold Spring Harbor Laboratory Press |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT hollensenannekruse improvedmicrornasuppressionbywprelinkedtoughdecoymicrornasponges AT thomsenrune improvedmicrornasuppressionbywprelinkedtoughdecoymicrornasponges AT bakrasmuso improvedmicrornasuppressionbywprelinkedtoughdecoymicrornasponges AT petersencharlottechristie improvedmicrornasuppressionbywprelinkedtoughdecoymicrornasponges AT ermegaardevar improvedmicrornasuppressionbywprelinkedtoughdecoymicrornasponges AT aagaardlars improvedmicrornasuppressionbywprelinkedtoughdecoymicrornasponges AT damgaardchristiankroun improvedmicrornasuppressionbywprelinkedtoughdecoymicrornasponges AT mikkelsenjacobgiehm improvedmicrornasuppressionbywprelinkedtoughdecoymicrornasponges |