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PMID: 17592038 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Isolation of microRNA targets by miRNP immunopurification.

RNA (New York, N.Y.) ·Vol. 13 ·No. 8 ·2007-08-00 ·Pages 1198-204

Easow G, Teleman AA, Cohen SM

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.

MeSH Terms
Animals Argonaute Proteins Drosophila/cytology,genetics,metabolism Drosophila Proteins/metabolism Eukaryotic Initiation Factors Immunoprecipitation/methods MicroRNAs/metabolism Mutation Ribonucleoproteins/isolation & purification,metabolism
Chemicals
AGO1 protein, Drosophila Argonaute Proteins Drosophila Proteins Eukaryotic Initiation Factors MicroRNAs Ribonucleoproteins
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Easow George
European Molecular Biology Laboratory, Heidelberg, Germany.
Teleman Aurelio A
Cohen Stephen M
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Article Info
Journal
RNA (New York, N.Y.)
Abbr.
RNA
ISSN
1355-8382
DOI
10.1261/rna.563707
Published
2007-08-00
Epub
2007-00-25
Pages
1198-204
Language
English
Region
United States
NLM ID
9509184
PMCID
PMC1924889
Subset
IM
External Links
PubMed source DOI: 10.1261/rna.563707 PMC Full Text View journal details
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