PT  - JOURNAL ARTICLE
AU  - Sarah E. Ochmann
AU  - Himanshu Joshi
AU  - Ece Büber
AU  - Henri G. Franquelim
AU  - Pierre Stegemann
AU  - Barbara Saccà
AU  - Ulrich F. Keyser
AU  - Aleksei Aksimentiev
AU  - Philip Tinnefeld
TI  - DNA Origami Voltage Sensors for Transmembrane Potentials with Single-Molecule Sensitivity
AID  - 10.1101/2021.08.18.456762
DP  - 2021 Jan 01
TA  - bioRxiv
PG  - 2021.08.18.456762
4099  - http://biorxiv.org/content/early/2021/08/18/2021.08.18.456762.short
4100  - http://biorxiv.org/content/early/2021/08/18/2021.08.18.456762.full
AB  - Signal transmission in neurons goes along with changes in the transmembrane potential. To report them, different approaches including optical voltage-sensing dyes and genetically encoded voltage indicators have evolved. Here, we present a DNA nanotechnology-based system. Using DNA origami, we incorporate and optimize different properties such as membrane targeting and voltage sensing modularly. As a sensing unit, we use a hydrophobic red dye anchored to the membrane and an anionic green dye at the DNA connecting the DNA origami and the membrane dye anchor. Voltage-induced displacement of the anionic donor unit is read out by changes of Fluorescence Resonance Energy Transfer (FRET) of single sensors attached to liposomes. They show a FRET change of ∼5% for ΔΨ=100 mV and allow adapting the potential range of highest sensitivity. Further, the working mechanism is rationalized by molecular dynamics simulations. Our approach holds potential for the application as non-genetically encoded sensors at membranes.Competing Interest StatementThe authors have declared no competing interest.GEVIGenetically Encoded Voltage Indicator;FRETFluorescence Resonance Energy Transfer;ssDNAsingle-stranded DNA;MDMolecular Dynamic;TIRFTotal Internal Reflection Fluorescence;dsDNAdouble-stranded DNA;smFRETsingle-molecule FRET;ALExAlternating Laser Excitation;PRProximity Ratio;SEMStandard Error of the Mean;REUSReplica Exchange Umbrella Sampling.