A general approach to engineer positive-going eFRET voltage indicators

Ahmed S. Abdelfattah (), Rosario Valenti, Jihong Zheng, Allan Wong, Kaspar Podgorski, Minoru Koyama, Douglas S. Kim and Eric R. Schreiter ()
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Ahmed S. Abdelfattah: Howard Hughes Medical Institute
Rosario Valenti: Howard Hughes Medical Institute
Jihong Zheng: Howard Hughes Medical Institute
Allan Wong: Howard Hughes Medical Institute
Kaspar Podgorski: Howard Hughes Medical Institute
Minoru Koyama: Howard Hughes Medical Institute
Douglas S. Kim: Howard Hughes Medical Institute
Eric R. Schreiter: Howard Hughes Medical Institute

Nature Communications, 2020, vol. 11, issue 1, 1-8

Abstract: Abstract Imaging membrane voltage from genetically defined cells offers the unique ability to report spatial and temporal dynamics of electrical signaling at cellular and circuit levels. Here, we present a general approach to engineer electrochromic fluorescence resonance energy transfer (eFRET) genetically encoded voltage indicators (GEVIs) with positive-going fluorescence response to membrane depolarization through rational manipulation of the native proton transport pathway in microbial rhodopsins. We transform the state-of-the-art eFRET GEVI Voltron into Positron, with kinetics and sensitivity equivalent to Voltron but flipped fluorescence signal polarity. We further apply this general approach to GEVIs containing different voltage sensitive rhodopsin domains and various fluorescent dye and fluorescent protein reporters.

Date: 2020
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DOI: 10.1038/s41467-020-17322-1

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