Characterizing 3D RNA structure by single molecule FRET

Methods. 2016 Jul 1:103:57-67. doi: 10.1016/j.ymeth.2016.02.004. Epub 2016 Feb 4.

Abstract

The importance of elucidating the three dimensional structures of RNA molecules is becoming increasingly clear. However, traditional protein structural techniques such as NMR and X-ray crystallography have several important drawbacks when probing long RNA molecules. Single molecule Förster resonance energy transfer (smFRET) has emerged as a useful alternative as it allows native sequences to be probed in physiological conditions and allows multiple conformations to be probed simultaneously. This review serves to describe the method of generating a three dimensional RNA structure from smFRET data from the biochemical probing of the secondary structure to the computational refinement of the final model.

Keywords: 3D model; FRET; Fluorescence; RNA structure; Single-molecule.

Publication types

  • Review
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Fluorescence Polarization
  • Fluorescence Resonance Energy Transfer*
  • Fluorescent Dyes / chemistry
  • Models, Molecular
  • Nucleic Acid Conformation
  • RNA / chemistry*
  • RNA / ultrastructure
  • Staining and Labeling

Substances

  • Fluorescent Dyes
  • RNA