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Cold Spring Harbor Laboratory Press, RNA, 10(14), p. 2254-2259, 2008

DOI: 10.1261/rna.1133808

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Site-specific crosslinking of human microRNPs to RNA targets

Journal article published in 2008 by Yohei Kirino ORCID, Zissimos Mourelatos ORCID
This paper is made freely available by the publisher.
This paper is made freely available by the publisher.

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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.