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


Steffen, Fabio D; Börner, Richard; Freisinger, Eva; Sigel, Roland K O (2019). Stick, Flick, Click: DNA-guided Fluorescent Labeling of Long RNA for Single-molecule FRET. CHIMIA International Journal for Chemistry, 73(4):257-261.

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.

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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Additional indexing

Item Type:Journal Article, refereed, original work
Communities & Collections:07 Faculty of Science > Department of Chemistry
Dewey Decimal Classification:540 Chemistry
Uncontrolled Keywords:Bioorthogonal · Fluorescence · Nucleic acids · Riboswitch · Spectroscopy
Language:English
Date:24 April 2019
Deposited On:29 May 2019 12:05
Last Modified:29 May 2019 12:05
Publisher:Swiss Chemical Society
ISSN:0009-4293
OA Status:Closed
Free access at:Publisher DOI. An embargo period may apply.
Publisher DOI:https://doi.org/10.2533/chimia.2019.257
Project Information:
  • : FunderUniversität Zürich
  • : Grant IDFK-17-098
  • : Project Title
  • : FunderUniversität Zürich
  • : Grant IDFK-14-096
  • : Project Title
  • : FunderUniversität Zürich
  • : Grant IDFK-15-095
  • : Project Title
  • : FunderSystemsX
  • : Grant ID
  • : Project Title
  • : FunderSNF
  • : Grant ID
  • : Project Title

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Embargo till: 2020-04-01