Inhibiting pri-miRNA processing with target site blockers

Annita Louloupi, Ulf Andersson Vang Ørom*

*Corresponding author af dette arbejde

Publikation: Bidrag til bog/antologi/rapport/proceedingBidrag til bog/antologiForskningpeer review

Abstract

The pri-miRNA processing is important for the final regulatory role of miRNAs on the expression of their target transcripts. The processing variability between pri-miRNAs can determine the final miRNA abundance better than primary transcription itself. Thus studying the in vivo pri-miRNA biogenesis could give more insights into the contribution of each individual miRNA on regulation of gene expression. Interfering processing of a specific pri-miRNA has been challenging due to the nature of the current RNA interfence methods. Here, we describe step by step a method to arrest processing of specific pri-miRNAs in vivo using LNA microRNA Target Site Blockers. We explain in detail the various aspects of this approach that can easily be applied to different mammalian cell types. The nature of this protocol allows the in vivo study of pri-miRNA processing and processing kinetics in cells treated with different conditions, mutants, and/or cancer cell lines under physiological conditions.

OriginalsprogEngelsk
TitelmiRNA Biogenesis
RedaktørerU. Ørum
Antal sider6
ForlagSpringer
Publikationsdatojan. 2018
Sider63-68
ISBN (Trykt)978-1-4939-8623-1
ISBN (Elektronisk)978-1-4939-8624-8
DOI
StatusUdgivet - jan. 2018
NavnMethods in Molecular Biology
Vol/bind1823
ISSN1064-3745

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