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Microprocessor dynamics shows co- and post-transcriptional processing of pri-miRNAs

miRNAs are small regulatory RNAs involved in the regulation of translation of target transcripts. miRNA biogenesis is a multistep process starting with the cleavage of the primary miRNA transcript in the nucleus by the Microprocessor complex. Endogenous processing of pri-miRNAs is challenging to stu...

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Detalles Bibliográficos
Autores principales: Louloupi, Annita, Ntini, Evgenia, Liz, Julia, Ørom, Ulf Andersson
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/PMC5435862/
https://www.ncbi.nlm.nih.gov/pubmed/28250203
http://dx.doi.org/10.1261/rna.060715.117
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author Louloupi, Annita
Ntini, Evgenia
Liz, Julia
Ørom, Ulf Andersson
author_facet Louloupi, Annita
Ntini, Evgenia
Liz, Julia
Ørom, Ulf Andersson
author_sort Louloupi, Annita
collection PubMed
description miRNAs are small regulatory RNAs involved in the regulation of translation of target transcripts. miRNA biogenesis is a multistep process starting with the cleavage of the primary miRNA transcript in the nucleus by the Microprocessor complex. Endogenous processing of pri-miRNAs is challenging to study and the in vivo kinetics of this process is not known. Here, we present a method for determining the processing kinetics of pri-miRNAs within intact cells over time, using a pulse-chase approach to label transcribed RNA during 15 min, and follow the processing within a 1-hour window after labeling with bromouridine. We show that pri-miRNAs exhibit different processing kinetics ranging from fast over intermediate to slow processing, and we provide evidence that pri-miRNA processing can occur both cotranscriptionally and post-transcriptionally.
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spelling pubmed-54358622017-06-02 Microprocessor dynamics shows co- and post-transcriptional processing of pri-miRNAs Louloupi, Annita Ntini, Evgenia Liz, Julia Ørom, Ulf Andersson RNA Article miRNAs are small regulatory RNAs involved in the regulation of translation of target transcripts. miRNA biogenesis is a multistep process starting with the cleavage of the primary miRNA transcript in the nucleus by the Microprocessor complex. Endogenous processing of pri-miRNAs is challenging to study and the in vivo kinetics of this process is not known. Here, we present a method for determining the processing kinetics of pri-miRNAs within intact cells over time, using a pulse-chase approach to label transcribed RNA during 15 min, and follow the processing within a 1-hour window after labeling with bromouridine. We show that pri-miRNAs exhibit different processing kinetics ranging from fast over intermediate to slow processing, and we provide evidence that pri-miRNA processing can occur both cotranscriptionally and post-transcriptionally. Cold Spring Harbor Laboratory Press 2017-06 /pmc/articles/PMC5435862/ /pubmed/28250203 http://dx.doi.org/10.1261/rna.060715.117 Text en © 2017 Louloupi et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society http://creativecommons.org/licenses/by-nc/4.0/ This article, published in RNA, 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
Louloupi, Annita
Ntini, Evgenia
Liz, Julia
Ørom, Ulf Andersson
Microprocessor dynamics shows co- and post-transcriptional processing of pri-miRNAs
title Microprocessor dynamics shows co- and post-transcriptional processing of pri-miRNAs
title_full Microprocessor dynamics shows co- and post-transcriptional processing of pri-miRNAs
title_fullStr Microprocessor dynamics shows co- and post-transcriptional processing of pri-miRNAs
title_full_unstemmed Microprocessor dynamics shows co- and post-transcriptional processing of pri-miRNAs
title_short Microprocessor dynamics shows co- and post-transcriptional processing of pri-miRNAs
title_sort microprocessor dynamics shows co- and post-transcriptional processing of pri-mirnas
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5435862/
https://www.ncbi.nlm.nih.gov/pubmed/28250203
http://dx.doi.org/10.1261/rna.060715.117
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