Optimization of PAR-CLIP for transcriptome-wide identification of binding sites of RNA-binding proteins.
Optimization of PAR-CLIP for transcriptome-wide identification of binding sites of RNA-binding proteins.
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DOI：
10.1016/j.ymeth.2016.10.007
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发表时间：
2017-04-15
期刊：
影响因子：
--
通讯作者：
Tuschl T
中科院分区：
文献类型：
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作者：
Garzia A;Meyer C;Morozov P;Sajek M;Tuschl T
可光激活核糖核苷增强的交联和免疫沉淀(PAR-CLIP)与下一代测序相结合是识别RNA结合蛋白(RBPs)内源性靶点的有效方法。根据每个RBP的特点，必须优化PAR-CLIP过程中的关键步骤。在这里，我们提出了一个全面的逐步PAR-CLIP协议，并详细解释了关键步骤。此外，我们报告了一种新的PAR-CLIP数据分析流水线应用于针对不同注释类别的细胞RNA的三个不同的RBP。
Photoactivatable-Ribonucleoside-Enhanced Crosslinking and Immunoprecipitation (PAR-CLIP) in combination with next-generation sequencing is a powerful method for identifying endogenous targets of RNA-binding proteins (RBPs). Depending on the characteristics of each RBP, key steps in the PAR-CLIP procedure must be optimized. Here we present a comprehensive step-by-step PAR-CLIP protocol with detailed explanations of the critical steps. Furthermore, we report the application of a new PAR-CLIP data analysis pipeline to three distinct RBPs targeting different annotation categories of cellular RNAs.