Homotransfer of FRET Reporters for Live Cell Imaging

Förster resonance energy transfer (FRET) between fluorophores of the same species was recognized in the early to mid-1900s, well before modern heterotransfer applications. Recently, homotransfer FRET principles have re-emerged in biosensors that incorporate genetically encoded fluorescent proteins....

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Bibliographic Details
Published in:Biosensors (Basel) 2018-10, Vol.8 (4), p.89
Main Authors: Snell, Nicole E, Rao, Vishnu P, Seckinger, Kendra M, Liang, Junyi, Leser, Jenna, Mancini, Allison E, Rizzo, M A
Format: Article
Language:English
Subjects:
Anisotropy
Biosensing Techniques - methods
biosensor
Biosensors
Chemical compounds
Clustering
Data collection
Design
Dimerization
Efficiency
Electric fields
Energy
Energy transfer
Fluorescence
Fluorescence polarization
Fluorescence resonance energy transfer
Fluorescence Resonance Energy Transfer - methods
fluorescent protein
Fluorophores
FRET
Genetic code
GFP
Green Fluorescent Proteins - chemistry
homotransfer
Lifetime
Light
Microscopy
Microscopy, Fluorescence
Modulators
Optical measuring instruments
Polarization
Proteins
Review
Sensors
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Summary:Förster resonance energy transfer (FRET) between fluorophores of the same species was recognized in the early to mid-1900s, well before modern heterotransfer applications. Recently, homotransfer FRET principles have re-emerged in biosensors that incorporate genetically encoded fluorescent proteins. Homotransfer offers distinct advantages over the standard heterotransfer FRET method, some of which are related to the use of fluorescence polarization microscopy to quantify FRET between two fluorophores of identical color. These include enhanced signal-to-noise, greater compatibility with other optical sensors and modulators, and new design strategies based upon the clustering or dimerization of singly-labeled sensors. Here, we discuss the theoretical basis for measuring homotransfer using polarization microscopy, procedures for data collection and processing, and we review the existing genetically-encoded homotransfer biosensors.
ISSN:2079-6374
2079-6374
DOI:10.3390/bios8040089