| Genome-wide Mapping of Cellular Protein–RNA Interactions Enabled by Chemical Crosslinking |
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| 基金项目： | supported by the National Natural Science Foundation of China(Grant No.31270838);the National Basic Research Foundation of China(Grant No.2014CB964900) |
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| 摘 要： | RNA–protein interactions influence many biological processes. Identifying the binding sites of RNA-binding proteins（RBPs） remains one of the most fundamental and important challenges to the studies of such interactions. Capturing RNA and RBPs via chemical crosslinking allows stringent purification procedures that significantly remove the non-specific RNA and protein interactions. Two major types of chemical crosslinking strategies have been developed to date, i.e., UV-enabled crosslinking and enzymatic mechanism-based covalent capture. In this review, we compare such strategies and their current applications, with an emphasis on the technologies themselves rather than the biology that has been revealed. We hope such methods could benefit broader audience and also urge for the development of new methods to study RNA RBP interactions.
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| 关 键 词： | 相互作用 化学交联 CRNA 细胞蛋白 组映射 全基因 RNA结合蛋白 启用 |
| 收稿时间： | 2014 Mar 9 |
Genome-wide Mapping of Cellular Protein-RNA Interactions Enabled by Chemical Crosslinking |
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| Authors: | Xiaoyu Li Jinghui Song Chengqi Yi |
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| Institution: | Xiaoyu Li(State Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, Beijing 100871, China)；Jinghui Song(State Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, Beijing 100871, China)；Chengqi Yi(State Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, Beijing 100871, China)； |
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| Abstract: | RNA-protein interactions influence many biological processes. Identifying the binding sites of RNA-binding proteins (RBPs) remains one of the most fundamental and important chal-lenges to the studies of such interactions. Capturing RNA and RBPs via chemical crosslinking allows stringent purification procedures that significantly remove the non-specific RNA and protein interactions. Two major types of chemical crosslinking strategies have been developed to date, i.e., UV-enabled crosslinking and enzymatic mechanism-based covalent capture. In this review, we com-pare such strategies and their current applications, with an emphasis on the technologies themselves rather than the biology that has been revealed. We hope such methods could benefit broader audi-ence and also urge for the development of new methods to study RNA RBP interactions. |
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| Keywords: | Protein RNA interactions High-throughput sequencing Crosslinking RNA-binding proteins Aza-IP miCLIP |
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