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 strat...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.展开更多
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).
文摘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.
