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RNA. 2007 Aug;13(8):1198-204. Epub 2007 Jun 25.

Isolation of microRNA targets by miRNP immunopurification.

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1
European Molecular Biology Laboratory, Heidelberg, Germany.

Abstract

microRNAs (miRNAs) serve as post-transcriptional regulators of gene expression, by guiding effector complexes (miRNPs) to target RNAs. Although considerable progress has been made in computational methods to identify miRNA targets, only a relatively limited assessment of their ability to function in vivo has been reported. Here we describe an alternative approach to miRNA target identification based on a biochemical method for purifying miRNP complexes with associated miRNAs and bound mRNA targets. Microarray analysis revealed a high degree of enrichment for miRNA complementary sites in the 3'UTRs of the miRNP-associated mRNAs. mRNAs specifically associated with an individual miRNA were identified by comparing the miRNP-associated mRNAs from wild-type flies and mutant flies lacking miR-1, and their regulation by the miRNA was validated. This approach provides a means to identify functional miRNA targets based on their physical interaction in vivo.

PMID:
17592038
PMCID:
PMC1924889
DOI:
10.1261/rna.563707
[Indexed for MEDLINE]
Free PMC Article
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