A large size-selective DNA nanopore with sensing applications

Rasmus P. Thomsen, Mette Galsgaard Malle, Anders Hauge Okholm, Swati Krishnan, Søren S.-R. Bohr, Rasmus Schøler Sørensen, Oliver Ries, Stefan Vogel, Friedrich C. Simmel, Nikos S. Hatzakis () and Jørgen Kjems ()
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Rasmus P. Thomsen: Aarhus University
Mette Galsgaard Malle: University of Copenhagen
Anders Hauge Okholm: Aarhus University
Swati Krishnan: Technische Universität München
Søren S.-R. Bohr: University of Copenhagen
Rasmus Schøler Sørensen: Aarhus University
Oliver Ries: University of Southern Denmark
Stefan Vogel: University of Southern Denmark
Friedrich C. Simmel: Technische Universität München
Nikos S. Hatzakis: University of Copenhagen
Jørgen Kjems: Aarhus University

Nature Communications, 2019, vol. 10, issue 1, 1-10

Abstract: Abstract Transmembrane nanostructures like ion channels and transporters perform key biological functions by controlling flow of molecules across lipid bilayers. Much work has gone into engineering artificial nanopores and applications in selective gating of molecules, label-free detection/sensing of biomolecules and DNA sequencing have shown promise. Here, we use DNA origami to create a synthetic 9 nm wide DNA nanopore, controlled by programmable, lipidated flaps and equipped with a size-selective gating system for the translocation of macromolecules. Successful assembly and insertion of the nanopore into lipid bilayers are validated by transmission electron microscopy (TEM), while selective translocation of cargo and the pore mechanosensitivity are studied using optical methods, including single-molecule, total internal reflection fluorescence (TIRF) microscopy. Size-specific cargo translocation and oligonucleotide-triggered opening of the pore are demonstrated showing that the DNA nanopore can function as a real-time detection system for external signals, offering potential for a variety of highly parallelized sensing applications.

Date: 2019
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DOI: 10.1038/s41467-019-13284-1

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