A general approach to engineer positive-going eFRET voltage indicators

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...

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Bibliographic Details
Published in:Nature communications 2020-07, Vol.11 (1), p.3444-3444, Article 3444
Main Authors: Abdelfattah, Ahmed S., Valenti, Rosario, Zheng, Jihong, Wong, Allan, Podgorski, Kaspar, Koyama, Minoru, Kim, Douglas S., Schreiter, Eric R.
Format: Article
Language:English
Subjects:
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631/1647/527/2047
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Action potential
Amino acids
Circuits
Depolarization
Electrochromism
Energy transfer
Engineers
Equilibrium
Fluorescence
Fluorescence resonance energy transfer
Fluorescent dyes
Fluorescent indicators
Genetic code
Genetic engineering
Humanities and Social Sciences
Membrane potential
Membranes
Microorganisms
multidisciplinary
Mutagenesis
Mutation
Polarity
Proteins
Rhodopsin
Science
Science (multidisciplinary)
Voltage
Voltage indicators
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Summary: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. Genetically encoded voltage indicators (GEVIs) allow visualisation of fast action potentials in neurons but most are bright at rest and dimmer during an action potential. Here, the authors engineer electrochromic FRET GEVIs with fast, bright and positive-going fluorescence signals for in vivo imaging.
ISSN:2041-1723
2041-1723
DOI:10.1038/s41467-020-17322-1