A general approach to engineer positive-going eFRET voltage indicators

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Nature Communications. 2020 Jul 10;11(1)

A general approach to engineer positive-going eFRET voltage indicators Schreiter LabPodgorski LabGENIEPrimary & iPS Cell CultureMolecular Genomics Viral Tools

Abdelfattah AS, Valenti R, Zheng J, Wong A, Podgorski K, Koyama M, Kim DS, Schreiter ER, GENIE Project

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Abstract

We engineered 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 transformed the state-of-the-art eFRET GEVI Voltron into Positron, with kinetics and sensitivity equivalent to Voltron but flipped fluorescence signal polarity. We further applied this general approach to GEVIs containing different voltage sensitive rhodopsin domains and various fluorescent dye and fluorescent protein reporters.