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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.

SUBMITTER: Hafner M 

PROVIDER: S-EPMC3508682 | biostudies-literature | 2012 Oct

REPOSITORIES: biostudies-literature

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Genome-wide identification of miRNA targets by PAR-CLIP.

Hafner Markus M   Lianoglou Steve S   Tuschl Thomas T   Betel Doron D  

Methods (San Diego, Calif.) 20120819 2

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-  ...[more]

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