RiPCA: An Assay for the Detection of RNA-Protein Interactions in Live Cells.
RiPCA: An Assay for the Detection of RNA-Protein Interactions in Live Cells.
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DOI：
10.1002/cpz1.358
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发表时间：
2022-03
期刊：
影响因子：
--
通讯作者：
Garner AL
中科院分区：
文献类型：
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作者：
Rosenblum SL;Garner AL
研究和调节RNA及其RNA结合蛋白（RBP）之间相互作用的兴趣日益增加，需要使能技术。现有的检测RNA-蛋白质相互作用（RPI）的方法通常限于生物化学或裂解后方法或基于细胞的方法，这些方法需要添加基于RNA的亲和标签，例如MS 2发夹，从而使它们无法用于检测小的或高度加工的RNA。利用基于生物正交化学和分裂荧光素酶的技术，我们开发了一种用于检测活细胞中RPIs的检测方法。本文详细介绍了RiPCA，RNA相互作用与蛋白质介导的互补分析的协议和设计考虑。基本方案1：用于检测RNA-蛋白质相互作用的细胞测定的设计和优化
Increasing interest in studying and modulating the interactions between RNAs and their RNA-binding proteins (RBPs) has borne the need for enabling technologies. Existing means of detecting RNA-protein interactions (RPIs) are often limited to biochemical or post-lysis methods or cell-based methods that require the addition of an RNA-based affinity tag, such as the MS2 hairpin, precluding them from use in detecting small or highly processed RNAs. Taking advantage of bioorthogonal chemistry- and split-luciferase-based technologies, we developed an assay for the detection of RPIs in live cells. This article details the protocol and design considerations for RiPCA, RNA interaction with Protein-mediated Complementation Assay. Basic Protocol 1: Design and optimization of a cellular assay for the detection of RNA-protein interactions