A Covalent Approach for Site-Specific RNA Labeling in Mammalian Cells
A Covalent Approach for Site-Specific RNA Labeling in Mammalian Cells
复制标题
哺乳动物细胞中位点特异性 RNA 标记的共价方法
DOI：
10.1002/anie.201410433
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发表时间：
2015-04-07
影响因子：
16.6
通讯作者：
Wang, Jiangyun
中科院分区：
文献类型：
--
作者：
Li, Fahui;Dong, Jianshu;Wang, Jiangyun
RNA研究和RNA纳米技术的进展取决于通过体外和哺乳动物细胞中的化学方法以高精度操纵和探测RNA的能力。然而，共价RNA标记方法与目前的蛋白质标记策略的范围和通用性相媲美，是欠发达的。报道了一种利用tRNA（Ile 2）-胍基吡啶合成酶（Tias）和点击化学对哺乳动物细胞中RNA进行位点特异性和序列特异性共价标记的方法。的晶体结构的Tias在复杂的叠氮化物轴承胍丁胺类似物解决解开Tias/基板识别的结构基础。独特的RNA序列特异性和可塑性Tias/底物识别使得叠氮化物/炔基团能够在体外和哺乳动物细胞中位点特异性转移至感兴趣的RNA分子。随后的点击化学反应促进靶RNA的通用标记、功能化和可视化。
Advances in RNA research and RNA nanotechnology depend on the ability to manipulate and probe RNA with high precision through chemical approaches, both invitro and in mammalian cells. However, covalent RNA labeling methods with scope and versatility comparable to those of current protein labeling strategies are underdeveloped. A method is reported for the site- and sequence-specific covalent labeling of RNAs in mammalian cells by using tRNA(Ile2)-agmatidine synthetase (Tias) and click chemistry. The crystal structure of Tias in complex with an azide-bearing agmatine analogue was solved to unravel the structural basis for Tias/substrate recognition. The unique RNA sequence specificity and plastic Tias/substrate recognition enable the site-specific transfer of azide/alkyne groups to an RNA molecule of interest invitro and in mammalian cells. Subsequent click chemistry reactions facilitate the versatile labeling, functionalization, and visualization of target RNA.