核糖核酸
RNA结合蛋白
计算生物学
生物
基因
免疫沉淀
核苷酸
遗传学
基因表达
功能(生物学)
互补DNA
作者
Ina Huppertz,Jan Attig,Andrea D’Ambrogio,Laura E. Easton,Christopher R. Sibley,Yoichiro Sugimoto,Mojca Tajnik,Julian König,Jernej Ule
出处
期刊:Methods
[Elsevier]
日期:2014-02-01
卷期号:65 (3): 274-287
被引量:377
标识
DOI:10.1016/j.ymeth.2013.10.011
摘要
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.
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