Author/Editor		Huppertz, Ina; Attig, Jan; D'Ambrogio, Andrea; Easton, Laura; Sibley, Christopher R.; Sugimoto, Yoichiro; Tajnik, Mojca; König, Julian; Ule, Jernej
Title		iCLIP
Type		članek
Publication year		2013
ISSN		1046-2023 - Methods (San Diego, Calif.)
Language		eng
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 proteinRNA 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 proteinRNA 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 RNA beljakovine iCLIP RNA proteins