eprintid: 10044779 rev_number: 21 eprint_status: archive userid: 608 dir: disk0/10/04/47/79 datestamp: 2018-03-07 14:35:02 lastmod: 2021-09-25 23:36:05 status_changed: 2018-03-07 14:35:02 type: article metadata_visibility: show creators_name: Lee, FCY creators_name: Ule, J title: Advances in CLIP Technologies for Studies of Protein-RNA Interactions ispublished: pub divisions: UCL divisions: B02 divisions: C07 divisions: D07 divisions: F85 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 note: 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. 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. date: 2018-02-01 date_type: published publisher: CELL PRESS official_url: http://doi.org/10.1016/j.molcel.2018.01.005 oa_status: green full_text_type: other language: eng primo: open primo_central: open_green article_type_text: Review verified: verified_manual elements_id: 1535707 doi: 10.1016/j.molcel.2018.01.005 lyricists_name: Lee, Flora lyricists_name: Ule, Jernej lyricists_id: FCYLE55 lyricists_id: JULEX61 actors_name: Ule, Jernej actors_id: JULEX61 actors_role: owner full_text_status: public publication: Molecular Cell volume: 69 number: 3 pagerange: 354-369 pages: 16 issn: 1097-4164 citation: 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 <https://doi.org/10.1016/j.molcel.2018.01.005>. Green open access document_url: https://discovery.ucl.ac.uk/id/eprint/10044779/1/combined%20file.pdf