Ule, J;
Hwang, H-W;
Darnell, RB;
(2018)
The Future of Cross-Linking and Immunoprecipitation (CLIP).
Cold Spring Harbor Perspectives in Biology
, 10
(8)
, Article a032243. 10.1101/cshperspect.a032243.
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Abstract
To understand the assembly and functional outcomes of protein–RNA regulation, it is crucial to precisely identify the positions of such interactions. Cross-linking and immunoprecipitation (CLIP) serves this purpose by exploiting covalent protein–RNA cross-linking and RNA fragmentation, along with a series of stringent purification and quality control steps to prepare complementary DNA (cDNA) libraries for sequencing. Here we describe the core steps of CLIP, its primary variations, and the approaches to data analysis. We present the application of CLIP to studies of specific cell types in genetically engineered mice and discuss the mechanistic and physiologic insights that have already been gained from studies using CLIP. We conclude by discussing the future opportunities for CLIP, including studies of human postmortem tissues from disease patients and controls, RNA epigenetic modifications, and RNA structure. These and other applications of CLIP will continue to unravel fundamental gene regulatory mechanisms while providing important biologic and clinically relevant insights.
Type: | Article |
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Title: | The Future of Cross-Linking and Immunoprecipitation (CLIP) |
Open access status: | An open access version is available from UCL Discovery |
DOI: | 10.1101/cshperspect.a032243 |
Additional information: | This version is the author accepted manuscript. For information on re-use, please refer to the publisher’s terms and conditions. |
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/10056241 |
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