首页|Development of a Universal RNA Dual-Terminal Labeling Method for Sensing RNA-Ligand Interactions
Development of a Universal RNA Dual-Terminal Labeling Method for Sensing RNA-Ligand Interactions
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Dual labeling of an RNA can provide Förster resonance energy transfer(FRET)sensors for studying RNA fold-ing,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 effi-ciency.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 hair-pin 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.
dual labelingFörster resonance energy transfer sensorMichael cycloadditiontetrazine-cycloalkyne ligationRNA-ligand interaction
Longhuai Cheng、Dejun Ma、Jie Zhang、Xueying Kang、Yang Wu、Yi Zhao、Long Yi、Zhen Xi
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State Key Laboratory of Elemento-Organic Chemistry,Department of Chemical Biology,College of Chemistry,National Pesticide Engineering Research Center,Collaborative Innovation Center of Chemical Science and Engineer-ing,Nankai University,Tianjin 300071
State Key Laboratory of Organic-Inorganic Composites and Beijing Key Laboratory of Bioprocess,Beijing University of Chemical Technology(BUCT),Beijing 100029
Key Laboratory of Intelligent Information Processing,Research Center for Ubiquitous Computing Systems,Institute of Computing Technology,Chinese Academy of Sciences,Beijing 100190
NETL
NSTL
万方数据
维普
