Molecular transport through large-diameter DNA nanopores

Abstract

DNA-based nanopores are synthetic biomolecular membrane pores, whose geometry and chemical functionality can be tuned using the tools of DNA nanotechnology, making them promising molecular devices for applications in single-molecule biosensing and synthetic biology. Here we introduce a large DNA membrane channel with an ≈4 nm diameter pore, which has stable electrical properties and spontaneously inserts into flat lipid bilayer membranes. Membrane incorporation is facilitated by a large number of hydrophobic functionalizations or, alternatively, streptavidin linkages between biotinylated channels and lipids. The channel displays an Ohmic conductance of ≈3 nS, consistent with its size, and allows electrically driven translocation of single-stranded and double-stranded DNA analytes. Using confocal microscopy and a dye influx assay, we demonstrate the spontaneous formation of membrane pores in giant unilamellar vesicles. Pores can be created both in an outside-in and an inside-out configuration.

Evolution of Structural DNA Nanotechnology

“…For example, DNA origami designs have been utilized as smart lids or gatekeepers on the solid‐state nanopores, which could be used in many sensing and sequencing applications . Recently, these strategies are adopted for creating versatile and modular membrane‐based DNA nanopores …”

Biomimetic Compartments Scaffolded by Nucleic Acid Nanostructures

“…The Keyser group and collaborators designed a DNA transmembrane nanopore with a large pore‐area (6 nm in diameter) that resulted in a tenfold greater conductance than in previously reported artificial ion channels . A large‐diameter DNA nanopore, reported by the Simmel group, allowed the electrically driven translocation of ss‐ and dsDNA analytes . To regulate DNA nanopores, Göpfrich and co‐workers used voltage switch to open and close a DNA nanopore: a high voltage (≈100 mV) caused an electric‐field‐induced change of the DNA conformation and orientation thereby blocking the nanopore .…”

Artificial Signal Transduction across Membranes

“…Proteinengineering and nanopores. [135,136] Pores consisting of DNA can also be formed withoutr elying on the relatively large size of DNA origami. [82] B) Fusinga lamethicintoaleucine zipper createsam etal-gated ion channel.…”