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Sensors 2014, 14(7), 11691-11713; doi:10.3390/s140711691

Engineering Genetically Encoded FRET Sensors

Laboratory of Chemical Biology and Institute of Complex Molecular Systems (ICMS), Department of Biomedical Engineering, Eindhoven University of Technology, Eindhoven 5600MB, The Netherlands
Author to whom correspondence should be addressed.
Received: 19 May 2014 / Revised: 24 June 2014 / Accepted: 26 June 2014 / Published: 2 July 2014
(This article belongs to the Special Issue Intracellular Sensing)
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Förster Resonance Energy Transfer (FRET) between two fluorescent proteins can be exploited to create fully genetically encoded and thus subcellularly targetable sensors. FRET sensors report changes in energy transfer between a donor and an acceptor fluorescent protein that occur when an attached sensor domain undergoes a change in conformation in response to ligand binding. The design of sensitive FRET sensors remains challenging as there are few generally applicable design rules and each sensor must be optimized anew. In this review we discuss various strategies that address this shortcoming, including rational design approaches that exploit self-associating fluorescent domains and the directed evolution of FRET sensors using high-throughput screening.
Keywords: FRET; sensors; directed evolution; protein engineering; multiparameter imaging; fluorescent proteins FRET; sensors; directed evolution; protein engineering; multiparameter imaging; fluorescent proteins
This is an open access article distributed under the Creative Commons Attribution License (CC BY 3.0).

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Lindenburg, L.; Merkx, M. Engineering Genetically Encoded FRET Sensors. Sensors 2014, 14, 11691-11713.

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