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Inhibiting pri-miRNA processing with target site blockers

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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.

Original languageEnglish
Title of host publicationmiRNA Biogenesis
EditorsU. Ørum
Number of pages6
PublisherSpringer
Publication yearJan 2018
Pages63-68
ISBN (print)978-1-4939-8623-1
ISBN (Electronic)978-1-4939-8624-8
DOIs
Publication statusPublished - Jan 2018
SeriesMethods in Molecular Biology
Volume1823
ISSN1064-3745

    Research areas

  • BrU pulse-chase labeling, Pri-miRNA processing dynamics

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