期刊
SENSORS
卷 14, 期 7, 页码 11691-11713出版社
MDPI
DOI: 10.3390/s140711691
关键词
FRET; sensors; directed evolution; protein engineering; multiparameter imaging; fluorescent proteins
资金
- Netherlands Organization of Scientific Research (VIDI) [700.56.428]
- European Research Council [ERC-2011-StG280255]
Forster 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.
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