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:
影响因子:
--
通讯作者:
Hafner,Markus
中科院分区:
文献类型:
--
作者:
Benhalevy,Daniel;Hafner,Markus
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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