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

ABSTRACT: 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.

SUBMITTER: Louloupi A

PROVIDER: S-EPMC5435862 | biostudies-literature | 2017 Jun

REPOSITORIES: biostudies-literature

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

Louloupi Annita A Ntini Evgenia E Liz Julia J Ørom Ulf Andersson UA

RNA (New York, N.Y.) 20170301 6

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 ...[more]

PMID: 28250203

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