title: Genome-wide identification of miRNA targets by PAR-CLIP.
abstract: miRNAs are short (20-23 nt) RNAs that are loaded into proteins of the Argonaute (AGO) family and guide them to partially complementary target sites on mRNAs, resulting in mRNA destabilization and/or translational repression. It is estimated that about 60% of the mammalian genes are potentially regulated by miRNAs, and therefore methods for experimental miRNA target determination have become valuable tools for the characterization of posttranscriptional gene regulation. Here we present a step-by-step protocol and guidelines for the computational analysis for the large-scale identification of miRNA target sites in cultured cells by photoactivatable ribonucleoside enhanced crosslinking and immunoprecipitation (PAR-CLIP) of AGO proteins.
authors.affiliation: Laboratory of RNA Molecular Biology, Howard Hughes Medical Institute, The Rockefeller University, New York, NY, USA.
authors.forename: Markus
authors.initials: M
authors.lastname: Hafner
authors.forename: Steve
authors.initials: S
authors.lastname: Lianoglou
authors.forename: Thomas
authors.initials: T
authors.lastname: Tuschl
authors.forename: Doron
authors.initials: D
authors.lastname: Betel
mesh_terms: D000818:Animals; D060565:Argonaute Proteins / Q000737:chemistry / Q000235:genetics; D019295:Computational Biology / Q000379:methods; D005786:Gene Expression Regulation; D016678:Genome; D051379:Mice; D035683:MicroRNAs / Q000737:chemistry / Q000235:genetics; D012333:RNA, Messenger / Q000737:chemistry / Q000235:genetics; D012263:Ribonucleosides / Q000737:chemistry / Q000302:isolation & purification
chemical_list: D060565:Argonaute Proteins; D035683:MicroRNAs; D012333:RNA, Messenger; D012263:Ribonucleosides