DNA Origami Voltage Sensors for Transmembrane Potentials with Single-Molecule Sensitivity
DNA Origami Voltage Sensors for Transmembrane Potentials with Single-Molecule Sensitivity
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DOI：
10.1101/2021.08.18.456762
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发表时间：
2021-08
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通讯作者：
Sarah E. Ochmann;Himanshu Joshi;Ece Büber;Henri G. Franquelim;Pierre Stegemann;B. Saccà;U. Keyser;A. Aksimentiev;P. Tinnefeld
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作者：
Sarah E. Ochmann;Himanshu Joshi;Ece Büber;Henri G. Franquelim;Pierre Stegemann;B. Saccà;U. Keyser;A. Aksimentiev;P. Tinnefeld
神经元内的信号传递伴随着跨膜电位的变化。为了报告它们，不同的方法，包括光学电压传感染料和基因编码电压指示器已经发展。在这里，我们提出了一个基于DNA纳米技术的系统。利用DNA折纸技术，我们结合并优化了不同的特性，如膜靶向和电压传感模块。作为传感单元，我们将疏水红色染料固定在膜上，并在DNA上使用阴离子绿色染料连接DNA折纸和膜染料锚。电压诱导的阴离子供体单元位移是通过附着在脂质体上的单个传感器的荧光共振能量转移（FRET）的变化来读出的。当ΔΨ=100 mV时，它们显示FRET变化约5%，并允许调整最高灵敏度的电位范围。并通过分子动力学模拟对其工作机理进行了论证。我们的方法具有应用于膜上非遗传编码传感器的潜力。
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.