Lee, FCY;
Ule, J;
(2018)
Advances in CLIP Technologies for Studies of Protein-RNA Interactions.
Molecular Cell
, 69
(3)
pp. 354-369.
10.1016/j.molcel.2018.01.005.
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Abstract
RNA binding proteins (RBPs) regulate all aspects in the life cycle of RNA molecules. To elucidate the elements that guide RNA specificity, regulatory mechanisms, and functions of RBPs, methods that identify direct endogenous protein-RNA interactions are particularly valuable. UV crosslinking and immunoprecipitation (CLIP) purifies short RNA fragments that crosslink to a specific protein and then identifies these fragments by sequencing. When combined with high-throughput sequencing, CLIP can produce transcriptome-wide maps of RNA crosslink sites. The protocol is comprised of several dozen biochemical steps, and improvements made over the last 15 years have increased its resolution, sensitivity, and convenience. Adaptations of CLIP are also emerging in the epitranscriptomic field to map the positions of RNA modifications accurately. Here, we describe the rationale for each step in the protocol and discuss the impact of variations to help users determine the most suitable option.
| Type: | Article |
|---|---|
| Title: | Advances in CLIP Technologies for Studies of Protein-RNA Interactions |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.1016/j.molcel.2018.01.005 |
| Publisher version: | http://doi.org/10.1016/j.molcel.2018.01.005 |
| Language: | English |
| Additional information: | Crown Copyright © 2018 Published by Elsevier Inc. This version is the author accepted manuscript. For information on re-use, please refer to the publisher’s terms and conditions. |
| Keywords: | Science & Technology, Life Sciences & Biomedicine, Biochemistry & Molecular Biology, Cell Biology, TRANSCRIPTOME-WIDE IDENTIFICATION, SINGLE-NUCLEOTIDE-RESOLUTION, SMALL NUCLEAR RNAS, BINDING PROTEINS, HITS-CLIP, CROSS-LINKING, IN-VIVO, RIBONUCLEOPROTEIN COMPLEXES, MESSENGER-RNAS, TARGET SITES |
| 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/10044779 |
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