RT Journal Article
SR Electronic
T1 DNA Origami Voltage Sensors for Transmembrane Potentials with Single-Molecule Sensitivity
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2021.08.18.456762
DO 10.1101/2021.08.18.456762
A1 Sarah E. Ochmann
A1 Himanshu Joshi
A1 Ece Büber
A1 Henri G. Franquelim
A1 Pierre Stegemann
A1 Barbara Saccà
A1 Ulrich F. Keyser
A1 Aleksei Aksimentiev
A1 Philip Tinnefeld
YR 2021
UL http://biorxiv.org/content/early/2021/08/18/2021.08.18.456762.abstract
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.