Genome-wide identification of miRNA targets by PAR-CLIP.
| Autor: | Hafner M; Laboratory of RNA Molecular Biology, Howard Hughes Medical Institute, The Rockefeller University, New York, NY, USA., Lianoglou S, Tuschl T, Betel D |
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| Jazyk: | angličtina |
| Zdroj: | Methods (San Diego, Calif.) [Methods] 2012 Oct; Vol. 58 (2), pp. 94-105. Date of Electronic Publication: 2012 Aug 19. |
| DOI: | 10.1016/j.ymeth.2012.08.006 |
| Abstrakt: | 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. (Copyright © 2012 Elsevier Inc. All rights reserved.) |
| Databáze: | MEDLINE |
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