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Biosensor optimization using a FRET pair based on mScarlet red fluorescent protein and an mScarlet-derived green fluorescent protein

Khyati Gohil, Sheng-Yi Wu, Kei Takahashi-Yamashiro, View ORCID ProfileYi Shen, Robert E. Campbell
doi: https://doi.org/10.1101/2022.06.20.496847
Khyati Gohil
1Department of Chemistry, University of Alberta, Edmonton, AB, Canada
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Sheng-Yi Wu
1Department of Chemistry, University of Alberta, Edmonton, AB, Canada
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Kei Takahashi-Yamashiro
1Department of Chemistry, University of Alberta, Edmonton, AB, Canada
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Yi Shen
1Department of Chemistry, University of Alberta, Edmonton, AB, Canada
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  • For correspondence: yi.shen@ualberta.ca robert.e.campbell@ualberta.ca
Robert E. Campbell
1Department of Chemistry, University of Alberta, Edmonton, AB, Canada
2Department of Chemistry, The University of Tokyo, Tokyo, Japan
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  • For correspondence: yi.shen@ualberta.ca robert.e.campbell@ualberta.ca
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Abstract

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, Ca2+, 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.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-ND 4.0 International license.
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Posted June 20, 2022.
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Biosensor optimization using a FRET pair based on mScarlet red fluorescent protein and an mScarlet-derived green fluorescent protein
Khyati Gohil, Sheng-Yi Wu, Kei Takahashi-Yamashiro, Yi Shen, Robert E. Campbell
bioRxiv 2022.06.20.496847; doi: https://doi.org/10.1101/2022.06.20.496847
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Biosensor optimization using a FRET pair based on mScarlet red fluorescent protein and an mScarlet-derived green fluorescent protein
Khyati Gohil, Sheng-Yi Wu, Kei Takahashi-Yamashiro, Yi Shen, Robert E. Campbell
bioRxiv 2022.06.20.496847; doi: https://doi.org/10.1101/2022.06.20.496847

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