Aptamer-DNA Origami-Functionalized Solid-State Nanopores for Single-Molecule Sensing of G-Quadruplex Formation

We describe the single-molecule sensing of G-quadruplex (G-q) formation in target-bound aptamers using solid-state nanopores. In order to do this, we functionalize the solid-state nanopore using DNA origami sheets incorporated with the aptamer sequence. This provides a modular configuration for nanopore-based single-molecule sensors where changing the aptamer sequence can produce the required origami functionalization of the nanopore. Furthermore, the DNA origami sheet also contains a pore which can modulate the translocation dynamics of the target molecule through the nanopore. Using this system, we show that we can distinguish the formation of G-q formation in the well-studied thrombin aptamer.

Automated summary

We demonstrate the single-molecule sensing of G-quadruplex formation in target-bound aptamers using solid-state nanopores. By functionalizing the nanopores with DNA origami sheets containing the aptamer sequence, we provide a versatile configuration for nanopore-based single-molecule sensors. The DNA origami sheet also includes a pore that can modulate the translocation dynamics of the target molecule. Using this system, we successfully differentiate the formation of G-quadruplex in the well-studied thrombin aptamer.