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.
journal_name
Mol Celljournal_title
Molecular cellauthors
Lee FCY,Ule Jdoi
10.1016/j.molcel.2018.01.005subject
Has Abstractpub_date
2018-02-01 00:00:00pages
354-369issue
3eissn
1097-2765issn
1097-4164pii
S1097-2765(18)30005-4journal_volume
69pub_type
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