Site-specific crosslinking of human microRNPs to RNA targets
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.
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Footnotes
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Reprint requests to: Zissimos Mourelatos, Division of Neuropathology, Department of Pathology and Laboratory Medicine, University of Pennsylvania School of Medicine, 605A Stellar-Chance Building, 422 Curie Boulevard, Philadelphia, PA 19104, USA; e-mail: mourelaz{at}uphs.upenn.edu; fax: (215) 898-9969.
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Article published online ahead of print. Article and publication date are at http://www.rnajournal.org/cgi/doi/10.1261/rna.1133808.
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- Received April 11, 2008.
- Accepted June 20, 2008.
- Copyright © 2008 RNA Society
