Quick-irCLIP: Interrogating protein-RNA interactions using a rapid and simple cross-linking and immunoprecipitation technique
Quick-irCLIP: Interrogating protein-RNA interactions using a rapid and simple cross-linking and immunoprecipitation technique
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DOI：
10.1016/j.mex.2019.05.014
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发表时间：
2019-01-01
期刊：
影响因子：
1.9
通讯作者：
Farkas, Michael
中科院分区：
文献类型：
--
作者：
Kaczynski, Tadeusz;Hussain, Ali;Farkas, Michael
RNA 结合蛋白 (RBP) 在非编码 RNA 的生化加工和生理功能中发挥着重要作用。因此，随着人们对非编码 RNA 的兴趣不断扩大，完善能够探测蛋白质-RNA 相互作用的技术将在这些分子的表征方面变得更有价值。为了识别给定 RBP 结合的 RNA，交联和免疫沉淀 (CLIP) 及其迭代已被广泛使用，但这些方法可能很复杂、劳动密集型且耗时。在这里，我们描述了一种基于个体核苷酸分辨率 CLIP (iCLIP) 和红外 CLIP (irCLIP) 的快速且技术简单的方法。我们的方案称为 Quick-irCLIP，避免了令人困惑的步骤，可以在不到三天的时间内完成，并且能够以单核苷酸分辨率询问蛋白质-RNA 相互作用。 (C) 2019 作者。由 Elsevier B.V. 出版
RNA-binding proteins (RBPs) are instrumental in the biochemical processing and physiological functioning of non-coding RNAs. Therefore, as interest in non-coding RNAs continues to expand, refining the techniques capable of probing protein-RNA interactions will prove ever more valuable in the characterization of these molecules. To identify the RNAs bound by a given RBP, cross-linking and immunoprecipitation (CLIP) and its iterations have been widely utilized, but these approaches can be complex, labor-intensive, and time consuming. Here, we describe a rapid and technically simple method based upon individual nucleotide resolution CLIP (iCLIP) and infrared CLIP (irCLIP). Termed quick-irCLIP, our protocol circumvents confounding steps, can be completed in less than three days, and is capable of interrogating protein-RNA interactions at single nucleotide resolution. (C) 2019 The Author(s). Published by Elsevier B.V.