Proximity-CLIP Provides a Snapshot of Protein-Occupied RNA Elements at Subcellular Resolution and Transcriptome-Wide Scale.

Proximity-CLIP Provides a Snapshot of Protein-Occupied RNA Elements at Subcellular Resolution and Transcriptome-Wide Scale.
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Proximity-CLIP 提供亚细胞分辨率和转录组范围内蛋白质占据的 RNA 元素的快照。

DOI:
10.1007/978-1-0716-0712-1_17
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发表时间:
2020
期刊:
Methods in molecular biology (Clifton, N.J.)
影响因子:
--
通讯作者:
Hafner,Markus
Hafner,Markus
中科院分区:
--
文献类型:
--
作者:
Benhalevy,Daniel;Hafner,Markus

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在过去的二十年里,信使RNA(mRNA)在特定亚细胞位置的分布已经被研究。在技术上,RNA定位的研究落后于蛋白质定位。在这里,我们提供了一个详细的协议为邻近CLIP,我们的小组最近开发的一种方法,结合蛋白质的邻近生物素化与光活化核糖核苷增强的蛋白质-RNA交联,同时配置文件的蛋白质组,包括RNA结合蛋白(RBP)和RBP结合的转录组在任何给定的亚细胞区室。该方法是分馏独立的,也使研究本地化的RNA加工中间体,以及在细胞隔室特异性的方式被蛋白质占据的RNA上的调节顺式作用元件的识别。
The distribution of messenger RNAs (mRNAs) to specific subcellular locations has been studied for the past two decades. Technically, studies of RNA localization are lagging those related to protein localization. Here we provide a detailed protocol forProximity-CLIP, a method recently developed by our group, that combines proximity biotinylation of proteins with photoactivatable ribonucleoside-enhanced protein-RNA cross-linking to simultaneously profile the proteome including RNA-binding proteins (RBPs) and the RBP-bound transcriptome in any given subcellular compartment. The approach is fractionation independent and also enables studying localized RNA-processing intermediates, as well as the identification of regulatorycis-acting elements on RNAs occupied by proteins in a cellular compartment-specific manner.
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影响因子: 42.7
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