Stick, Flick, Click: DNA-guided Fluorescent Labeling of Long RNA for Single-molecule FRET

Authors

  • Fabio D. Steffen Department of Chemistry, University of Zurich, Winterthurerstrasse 190, CH-8057 Zurich, SCS-Metrohm Award for best oral presentation in Medicinal Chemistry & Chemical Biology;, Email: fabio.steffen@chem.uzh.ch
  • Richard Börner Department of Chemistry, University of Zurich, Winterthurerstrasse 190, CH-8057 Zurich
  • Eva Freisinger Department of Chemistry, University of Zurich, Winterthurerstrasse 190, CH-8057 Zurich
  • Roland K. O. Sigel Department of Chemistry, University of Zurich, Winterthurerstrasse 190, CH-8057 Zurich;, Email: roland.sigel@chem.uzh.ch

DOI:

https://doi.org/10.2533/chimia.2019.257

Keywords:

Bioorthogonal, Fluorescence, Nucleic acids, Riboswitch, Spectroscopy

Abstract

Exploring 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.

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Published

2019-04-24

How to Cite

[1]
F. D. Steffen, R. Börner, E. Freisinger, R. K. O. Sigel, Chimia 2019, 73, 257, DOI: 10.2533/chimia.2019.257.