Huppertz, I;
Attig, J;
D'Ambrogio, A;
Eastonb, L;
Sibley, CR;
Sugimoto, Y;
Tajnik, M;
... Ule, J; + view all
(2014)
iCLIP: Protein-RNA interactions at nucleotide resolution.
Methods
, 65
(3)
pp. 274-287.
10.1016/j.ymeth.2013.10.011.
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Abstract
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.
| Type: | Article |
|---|---|
| Title: | iCLIP: Protein-RNA interactions at nucleotide resolution. |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.1016/j.ymeth.2013.10.011 |
| Publisher version: | http://dx.doi.org/10.1016/j.ymeth.2013.10.011 |
| Additional information: | ©� 2013 The Authors. Published by Elsevier Inc. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/3.0/). |
| Keywords: | RNA, RNA-binding protein, UV crosslinking and immunoprecipitation (CLIP), high-throughput sequencing, iCLIP, post-transcriptional regulation, protein-RNA interaction |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences > UCL Queen Square Institute of Neurology UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences > UCL Queen Square Institute of Neurology > Department of Neuromuscular Diseases |
| URI: | https://discovery.ucl.ac.uk/id/eprint/1415709 |
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