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A large size-selective DNA nanopore with sensing applications

Rasmus P. ThomsenInterdisciplinary Nanoscience Center, Aarhus University, Aarhus C, 8000, DenmarkMette Galsgaard MalleDepartment of Chemistry & Nanoscience Center, University of Copenhagen, Universitetsparken 5, Copenhagen, 2100, DenmarkAnders H. OkholmArla Innovation Centre, Agro Food Park 19, 8200, Aarhus N, DenmarkSwati KrishnanPhysics Department and ZNN/WSI, Technische Universität München, 85748, Garching, GermanySøren S.-R. BohrDepartment of Chemistry & Nanoscience Center, University of Copenhagen, Universitetsparken 5, Copenhagen, 2100, DenmarkRasmus Schøler SørensenInterdisciplinary Nanoscience Center, Aarhus University, Aarhus C, 8000, DenmarkOliver RiesDepartment of Physics, Chemistry and Pharmacy, University of Southern Denmark, Campusvej 55, 5230, Odense M, DenmarkStefan VogelDepartment of Physics, Chemistry and Pharmacy, University of Southern Denmark, Campusvej 55, 5230, Odense M, DenmarkFriedrich C. SimmelPhysics Department and ZNN/WSI, Technische Universität München, 85748, Garching, GermanyNikos S. HatzakisDepartment of Chemistry & Nanoscience Center, University of Copenhagen, Universitetsparken 5, Copenhagen, 2100, Denmark. [email protected]Jørgen KjemsDepartment of Molecular Biology and Genetics, Aarhus University, Aarhus C, 8000, Denmark. [email protected]

2019en

ABI

Annotatsiya

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.

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Identifikatorlar

DOI: 10.1038/s41467-019-13284-1

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