A Covalent Approach for Site-Specific RNA Labeling in Mammalian Cells

• Published in: Angewandte Chemie (International ed.) Vol. 54; no. 15; pp. 4597 - 4602
• Main Authors: Li, Fahui, Dong, Jianshu, Hu, Xiaosong, Gong, Weimin, Li, Jiasong, Shen, Jing, Tian, Huifang, Wang, Jiangyun
• Format: Journal Article
• Language: English
• Published: Weinheim WILEY-VCH Verlag 07.04.2015; WILEY‐VCH Verlag; Wiley; Wiley Subscription Services, Inc
• Edition: International ed. in English
• Subjects: Alkynes; Alkynes - chemistry; Animals; Archaea - enzymology; Azides - chemistry; bioorthogonal chemistry; biotechnology; Cell Line; Chemistry; Chemistry, Multidisciplinary; Click Chemistry; Covalence; Crystal structure; Cytidine - analogs & derivatives; Cytidine - chemistry; Cytidine - metabolism; Fluorescent Dyes - chemistry; Humans; Labeling; Ligases - metabolism; Marking; Models, Molecular; Optical Imaging; Physical Sciences; Proteins; Recognition; Ribonucleic acids; RNA labeling; RNA modification; RNA, Transfer - analysis; RNA, Transfer - metabolism; Science & Technology; Staining and Labeling; Strategy; tRNA; Versatility; RNA modification; biotechnology; COMPLEX; tRNA; SINGLE-MOLECULE; CLICK CHEMISTRY; RNA labeling; bioorthogonal chemistry; LIVE CELLS; GENETIC-CODE; CYTIDINE; STRUCTURAL BASIS; PROTEINS; LIVING CELLS
• ISSN: 1433-7851; 1521-3773
• DOI: 10.1002/anie.201410433

Advances in RNA research and RNA nanotechnology depend on the ability to manipulate and probe RNA with high precision through chemical approaches, both in vitro 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 tRNAIle2‐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 in vitro and in mammalian cells. Subsequent click chemistry reactions facilitate the versatile labeling, functionalization, and visualization of target RNA. Tag and click: The ability to specifically label RNAs in vitro and in mammalian cells would be highly significant for RNA research, however, covalent RNA labeling methods with scope and versatility comparable to those for protein labeling have not been reported. A method was developed for the site‐ and sequence‐specific covalent labeling of RNAs in mammalian cells, based on the action of tRNAIle2‐agmatidine synthetase (Tias) and click chemistry.