Site-specific Crosslinking of Human microRNPs to RNA Targets

RNA. 2008 Oct;14(10):2254-9. doi: 10.1261/rna.1133808. Epub 2008 Aug 21.

Abstract

MicroRNAs (miRNAs) regulate the expression of numerous genes and are implicated in the pathogenesis of many human diseases. miRNAs act as specificity determinants to guide deposition of microRNPs (miRNPs) onto miRNA recognition elements (MREs) that are found in mRNA targets. We have adapted a site-specific crosslinking approach, previously used in the analysis of splicing, to interrogate protein factors that physically associate with MREs in human cells. We find that Ago proteins are the only proteins that bind to the MRE in a miRNA-dependent manner. Our method may be used in various experimental systems to analyze protein factors that influence MRE accessibility by miRNAs and the composition of MRE-bound miRNPs.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Humans
  • Methods
  • MicroRNAs / antagonists & inhibitors
  • MicroRNAs / chemistry*
  • MicroRNAs / radiation effects
  • Ribonucleoproteins / chemistry*
  • Ultraviolet Rays

Substances

  • MicroRNAs
  • Ribonucleoproteins
  • mirnlet7 microRNA, human