Genome-wide identification of miRNA targets by PAR-CLIP

被引：79

作者：

Hafner, Markus ^{[1
]}

Lianoglou, Steve ^{[2
,3
]}

Tuschl, Thomas ^{[1
]}

Betel, Doron ^{[4
,5
]}

机构：

[1] Rockefeller Univ, Howard Hughes Med Inst, Lab RNA Mol Biol, New York, NY 10021 USA

[2] Mem Sloan Kettering Canc Ctr, Computat Biol Program, New York, NY 10021 USA

[3] Weill Cornell Med Coll, Dept Physiol Biophys & Syst Biol, New York, NY USA

[4] Weill Cornell Med Coll, Dept Med, New York, NY USA

[5] Weill Cornell Med Coll, Inst Computat Biomed, New York, NY USA

来源：

METHODS | 2012年 / 58卷 / 02期

关键词：

CLIP; miRNA; Next-generation sequencing; Bioinformatics; RNA-BINDING PROTEIN; MICRORNA TARGETS; SITES; SEQUENCE; ALIGNMENT; INSIGHTS; REVEALS; BIOLOGY; CELLS; DNA;

D O I：

10.1016/j.ymeth.2012.08.006

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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. (C) 2012 Elsevier Inc. All rights reserved.

引用

页码：94 / 105

页数：12

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