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| Item Type: | Article |
|---|---|
| Title: | iCLIP: protein-RNA interactions at nucleotide resolution |
| Creators Name: | Huppertz, I. and Attig, J. and D'Ambrogio, A. and Easton, L.E. and Sibley, C.R. and Sugimoto, Y. and Tajnik, M. and König, J. and Ule, J. |
| 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. |
| Keywords: | iCLIP, UV Crosslinking and Immunoprecipitation (CLIP), Protein-RNA Interaction, High-Throughput Sequencing, RNA-Binding Protein, RNA, Post-Transcriptional Regulation |
| Source: | Methods |
| ISSN: | 1046-2023 |
| Publisher: | Elsevier |
| Volume: | 65 |
| Number: | 3 |
| Page Range: | 274-87 |
| Date: | February 2014 |
| Official Publication: | https://doi.org/10.1016/j.ymeth.2013.10.011 |
| PubMed: | View item in PubMed |
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