title: Engineering genetically encoded FRET sensors.
abstract: 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.
authors.affiliation: Laboratory of Chemical Biology and Institute of Complex Molecular Systems (ICMS), Department of Biomedical Engineering, Eindhoven University of Technology, Eindhoven 5600MB, The Netherlands. laurens.lindenburg@googlemail.com.
authors.forename: Laurens
authors.initials: L
authors.lastname: Lindenburg
authors.affiliation: Laboratory of Chemical Biology and Institute of Complex Molecular Systems (ICMS), Department of Biomedical Engineering, Eindhoven University of Technology, Eindhoven 5600MB, The Netherlands. m.merkx@tue.nl.
authors.forename: Maarten
authors.initials: M
authors.lastname: Merkx
mesh_terms: D000818:Animals; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D006801:Humans; D008164:Luminescent Proteins / Q000032:analysis* / Q000235:genetics*; D057054:Molecular Imaging / Q000379:methods*; D015202:Protein Engineering / Q000379:methods*; D025941:Protein Interaction Mapping / Q000379:methods*
chemical_list: D008164:Luminescent Proteins