Capturing microRNA targets using an RNA-induced silencing complex (RISC)-trap approach.

Abstract:

:Identifying targets is critical for understanding the biological effects of microRNA (miRNA) expression. The challenge lies in characterizing the cohort of targets for a specific miRNA, especially when targets are being actively down-regulated in miRNA- RNA-induced silencing complex (RISC)-messengerRNA (mRNA) complexes. We have developed a robust and versatile strategy called RISCtrap to stabilize and purify targets from this transient interaction. Its utility was demonstrated by determining specific high-confidence target datasets for miR-124, miR-132, and miR-181 that contained known and previously unknown transcripts. Two previously unknown miR-132 targets identified with RISCtrap, adaptor protein CT10 regulator of kinase 1 (CRK1) and tight junction-associated protein 1 (TJAP1), were shown to be endogenously regulated by miR-132 in adult mouse forebrain. The datasets, moreover, differed in the number of targets and in the types and frequency of microRNA recognition element (MRE) motifs, thus revealing a previously underappreciated level of specificity in the target sets regulated by individual miRNAs.

authors

Cambronne XA,Shen R,Auer PL,Goodman RH

doi

10.1073/pnas.1218887109

subject

Has Abstract

pub_date

2012-12-11 00:00:00

pages

20473-8

issue

50

eissn

0027-8424

issn

1091-6490

pii

1218887109

journal_volume

109

pub_type

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