Abstract / Description of output
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.
Original language | English |
---|---|
Pages (from-to) | 274-287 |
Number of pages | 14 |
Journal | Methods |
Volume | 65 |
Issue number | 3 |
DOIs | |
Publication status | Published - Feb 2014 |
Keywords / Materials (for Non-textual outputs)
- binding sites
- DNA
- Gene Expression Regulation
- gene library
- HeLa Cells
- High-Throughput Nucleotide Sequencing
- humans
- Immunoprecipitation/methods
- protein binding
- RNA/chemistry
- RNA-Binding Proteins/chemistry
- ultraviolet rays
- circular/chemistry