iCLIP: protein-RNA interactions at nucleotide resolution.
Methods
; 65(3): 274-87, 2014 Feb.
Article
in En
| MEDLINE
| ID: mdl-24184352
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.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
RNA
/
Gene Library
/
RNA-Binding Proteins
/
Immunoprecipitation
Limits:
Humans
Language:
En
Journal:
Methods
Journal subject:
BIOQUIMICA
Year:
2014
Document type:
Article
Affiliation country:
United kingdom