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通过 iCLIP 方法鉴定斑马鱼胚胎转录组中的 RNA 结合蛋白图谱。

Identification of RNA-Binding Protein Landscapes Across Zebrafish Embryonic Transcriptome via iCLIP Approach.

机构信息

Department of Pharmacology, Weill Cornell Medicine, New York, NY, USA.

出版信息

Methods Mol Biol. 2021;2218:331-345. doi: 10.1007/978-1-0716-0970-5_26.

Abstract

In metazoans, fertilization initiates vast remodeling of the embryonic proteome and transcriptome. This is accomplished via complex post-transcriptional regulation of maternal and zygotic RNA. RNA-binding proteins (RBPs) are one of the major mediators of embryonic post-transcriptional RNA regulation. Thus, elucidation of the molecular mechanisms by which maternal and zygotic transcripts change their translational capacities and expression levels requires thorough and precise determination of the targets and binding sites of individual RBPs in embryonic transcriptomes. Here, I provide a detailed protocol for the UV crosslinking-based method, named iCLIP, to study RBP functions during early zebrafish embryogenesis.

摘要

在后生动物中,受精启动了胚胎蛋白质组和转录组的广泛重构。这是通过母体和合子 RNA 的复杂转录后调控来实现的。RNA 结合蛋白(RBPs)是胚胎转录后 RNA 调控的主要介导者之一。因此,阐明母体和合子转录物如何改变其翻译能力和表达水平的分子机制,需要对胚胎转录组中单个 RBP 的靶标和结合位点进行彻底和精确的确定。在这里,我提供了一种基于 UV 交联的方法(称为 iCLIP)的详细方案,用于研究早期斑马鱼胚胎发生过程中 RBP 的功能。

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