Methods 2016 07 4;103:57-67. Epub 2016 Feb 4.
Department of Medicine, University of Cambridge, Addenbrooke's Hospital, Hills Rd, Cambridge CB2 0QQ, UK. Electronic address:
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.