Stick, Flick, Click: DNA-guided Fluorescent Labeling of Long RNA for Single-molecule FRET
Keywords:Bioorthogonal, Fluorescence, Nucleic acids, Riboswitch, Spectroscopy
AbstractExploring the spatiotemporal dynamics of biomolecules on a single-molecule level requires innovative ways to make them spectroscopically visible. Fluorescence resonance energy transfer (FRET) uses a pair of organic dyes as reporters to measure distances along a predefined biomolecular reaction coordinate. For this nanoscopic ruler to work, the fluorescent labels need to be coupled onto the molecule of interest in a bioorthogonal and site-selective manner. Tagging large non-coding RNAs with single-nucleotide precision is an open challenge. Here we summarize current strategies in labeling riboswitches and ribozymes for fluorescence spectroscopy and FRET in particular. A special focus lies on our recently developed, DNA-guided approach that inserts two fluorophores through a stepwise process of templated functionality transfer and click chemistry.
How to Cite
F. D. Steffen, R. Börner, E. Freisinger, R. K. O. Sigel, Chimia 2019, 73, 257, DOI: 10.2533/chimia.2019.257.
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