Development of a Universal RNA Dual-Terminal Labeling Method for Sensing RNA-Ligand Interactions

作     者：Longhuai Cheng Dejun Ma Jie Zhang Xueying Kang Yang Wu Yi Zhao Long Yi Zhen Xi Longhuai Cheng;Dejun Ma;Jie Zhang;Xueying Kang;Yang Wu;Yi Zhao;Long Yi;Zhen Xi

作者机构：State Key Laboratory of Elemento-Organic ChemistryDepartment of Chemical BiologyCollege of ChemistryNational Pesticide Engineering Research CenterCollaborative Innovation Center of Chemical Science and EngineeringNankai UniversityTianjin 300071 State Key Laboratory of Organic-Inorganic Composites and Beijing Key Laboratory of BioprocessBeijing University of Chemical Technology(BUCT)Beijing 100029 Key Laboratory of Intelligent Information ProcessingResearch Center for Ubiquitous Computing SystemsInstitute of Computing TechnologyChinese Academy of SciencesBeijing 100190 

出 版 物：《CCS Chemistry》 (中国化学会会刊（英文）)

年 卷 期：2023年第5卷第1期

页      面：221-233页

核心收录：

学科分类：08[工学] 080202[工学-机械电子工程] 0802[工学-机械工程] 0703[理学-化学] 

基　　金：This research was made possible as a result of a generous grant from the National Nature Science Foundation of China(grant nos.21877008,32070670,and 22177010) the Zhejiang Provincial Natural Science Foundation of China(grant no.LY21C060003). 

主　　题：dual labeling Förster resonance energy transfer sensor Michael cycloaddition tetrazinecycloalkyne ligation RNA-ligand interaction 

摘      要：Dual labeling of an RNA can provide Förster resonance energy transfer(FRET)sensors for studying RNA folding,miRNA maturation,and RNA-protein interactions.Here,we report the development of a highly efficient strategy for direct dual-terminal labeling of any RNA of interest.We explored new Michael cycloaddition for facile labeling of 5′-terminal RNA with improved efficiency.Direct chemical tetrazinylation of RNA at the 3′-terminus was achieved with the highly efficient and catalysis-free tetrazine-cycloalkyne ligation.Both single-terminal labeling methods were combined for dual-terminal labeling of an RNA including short hairpin RNA,pre-miRNA,riboswitch,and noncoding RNA.Notably,these dual-labeled RNA-based FRET sensors were used to monitor RNA-ligand interactions in vitro and in live cells.It is anticipated that these universal RNA labeling strategies will be useful to study RNA structures and functions.