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

Xiaolu A. Cambronne, Rongkun Shen, Paul L. Auer, Richard H. Goodman

Research output: Contribution to journalArticle

38 Scopus citations

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 highconfidence target datasets for miR-124, miR-132, and miR-181 that contained known and previously unknown transcripts. Two previously unknown miR-132 targets identifi ed 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.

Original languageEnglish (US)
Pages (from-to)20473-20478
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume109
Issue number50
DOIs
StatePublished - Dec 11 2012

Keywords

  • Argonaute
  • GW182

ASJC Scopus subject areas

  • General

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