CLIP-Seq to identify targets and interactions of RNA binding proteins and RNA modifying enzymes.
CLIP-Seq to identify targets and interactions of RNA binding proteins and RNA modifying enzymes.
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DOI：
10.1016/bs.mie.2021.08.001
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发表时间：
2021
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RNA的化学修饰是目前发展最快的领域之一。在不同的RNA物种中，许多类型的RNA修饰已被证明在广泛的细胞过程中发挥多方面的作用。这些修改分别由writer和eraser酶安装和擦除。此外，RNA化学修饰通过影响RNA分子的化学或结构变化或通过识别修饰而具有下游生物学效应;这些功能主要由修饰阅读器蛋白执行。阅读器蛋白可以结合到修饰位点并引起下游信号级联。研究擦除器、写入器和读取器的基本工具之一是交联免疫沉淀，然后是高通量测序（CLIP-seq）。这种方法可以检测内源RNA上与RNA结合蛋白或RNA修饰酶结合的位点。基本上，这种策略允许细胞内发生的表转录组和分子事件的快照。在这篇文章中，我们详细介绍了CLIP测序中涉及的各个步骤。
The study of RNA chemical modifications is currently one of the rapid-growing fields. Many types of RNA modifications in diverse RNA species have been shown to play versatile roles in a wide array of cellular processes. These modifications are installed and erased by writer and eraser enzymes, respectively. Additionally, RNA chemical modifications have downstream biological effects through either influencing changes in the chemistry or structure of RNA molecules or through recognition of the modification; these functions are primarily executed by the modification reader proteins. Reader proteins may bind to the modification site and cause a downstream signal cascade. One of the essential tools for studying erasers, writers, and readers is cross-linking immunoprecipitation followed by high-throughput sequencing (CLIP-seq). This method can detect the sites on endogenous RNAs bound by RNA-binding proteins or RNA modifying enzymes. Essentially, this strategy allows for snapshots of the epitranscriptome and molecular events occurring within the cell. In this article, we go through in detail the various steps involved in CLIP-sequencing.
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