Biosensor Optimization Using a Förster Resonance Energy Transfer Pair Based on mScarlet Red Fluorescent Protein and an mScarlet-Derived Green Fluorescent Protein

Genetically encoded biosensors based on Förster resonance energy transfer (FRET) are indispensable tools for monitoring biochemical changes in cells. Green and red fluorescent protein-based FRET pairs offer advantages over the classically employed cyan and yellow fluorescent protein pairs, such as b...

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Bibliographic Details
Published in:ACS sensors 2023-02, Vol.8 (2), p.587-597
Main Authors: Gohil, Khyati, Wu, Sheng-Yi, Takahashi-Yamashiro, Kei, Shen, Yi, Campbell, Robert E
Format: Article
Language:English
Subjects:
Biosensing Techniques
Fluorescence Resonance Energy Transfer
Green Fluorescent Proteins - chemistry
Luminescent Proteins - chemistry
Red Fluorescent Protein
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Summary:Genetically encoded biosensors based on Förster resonance energy transfer (FRET) are indispensable tools for monitoring biochemical changes in cells. Green and red fluorescent protein-based FRET pairs offer advantages over the classically employed cyan and yellow fluorescent protein pairs, such as better spectral separation, lower phototoxicity, and less autofluorescence. Here, we describe the development of an mScarlet-derived green fluorescent protein (designated as mWatermelon) and its use as a FRET donor to the red fluorescent protein mScarlet-I as a FRET acceptor. We tested the functionality of this FRET pair by engineering biosensors for the detection of protease activity, Ca , and K . Furthermore, we described a strategy to enhance the FRET efficiency of these biosensors by modulating the intramolecular association between mWatermelon and mScarlet-I.
ISSN:2379-3694
2379-3694
DOI:10.1021/acssensors.2c01730