iCLIP: Protein-RNA interactions at nucleotide resolution

被引：308

作者：

Huppertz, Ina ^{[1
,2
]}

Attig, Jan ^{[1
,2
]}

D'Ambrogio, Andrea ^{[1
,2
]}

Easton, Laura E. ^{[2
]}

Sibley, Christopher R. ^{[1
,2
]}

Sugimoto, Yoichiro ^{[2
]}

Tajnik, Mojca ^{[2
,3
]}

Koenig, Julian ^{[1
,2
,4
]}

Ule, Jernej ^{[1
,2
]}

机构：

[1] UCL Inst Neurol, Dept Mol Neurosci, London WC1N 3BG, England

[2] MRC Lab Mol Biol, Cambridge CB2 0QH, England

[3] Univ Ljubljana, Fac Med, Ljubljana 1000, Slovenia

[4] Inst Mol Biol, D-55128 Mainz, Germany

来源：

METHODS | 2014年 / 65卷 / 03期

基金：

欧洲研究理事会; 英国医学研究理事会;

关键词：

iCLIP; UV crosslinking and immunoprecipitation (CLIP); Protein-RNA interaction; High-throughput sequencing; RNA-binding protein; RNA; Post-transcriptional regulation; BINDING PROTEINS; TRANSCRIPTOME; IDENTIFICATION; REVEALS; BIOLOGY; SITES; CLIP;

D O I：

10.1016/j.ymeth.2013.10.011

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

RNA-binding proteins (RBPs) are key players in the post-transcriptional regulation of gene expression. Precise knowledge about their binding sites is therefore critical to unravel their molecular function and to understand their role in development and disease. Individual-nucleotide resolution UV crosslinking and immunoprecipitation (iCLIP) identifies protein-RNA crosslink sites on a genome-wide scale. The high resolution and specificity of this method are achieved by an intramolecular cDNA circularization step that enables analysis of cDNAs that truncated at the protein-RNA crosslink sites. Here, we describe the improved iCLIP protocol and discuss critical optimization and control experiments that are required when applying the method to new RBPs. (C) 2013 The Authors. Published by Elsevier Inc.

引用

页码：274 / 287

页数：14

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