Towards the Therapeutic Application of DNA Origami

Abstract

DNA nanotechnology enables the construction of arbitrarily-shaped, complex and programmable nanostructures. By leveraging the self-assembly of a long single-stranded DNA scaffold with short single stranded DNA staples through Watson-Crick base pairing, DNA origami is an efficient and versatile process for creating precise nanoscale architectures. This programmability enables many potential applications of DNA origami nanostructures in multiple fields from materials to medicine. In particular, DNA origami show fundamental characteristics required of drug delivery systems: high biocompatibility and nanoscale control over size, shape, and surface functionalization. These are key advantages over clinical nanoparticles today such as liposomes or polymeric nanoparticles whose surfaces are typically homogenous. Further, DNA origami enable logic computation and already have shown incredible function ex-vivo as molecular machines. Here, we address key challenges in the design and implementation of therapeutic DNA origami. First, we determine the governing principles for modulation of cellular uptake of DNA origami by mammalian cells using shape and size. We next develop a novel method for extending the half-life of DNA origami in vivo, and demonstrate unprecedented survival of over 14 days under strenuous nuclease conditions, compared to <1 min for untreated structures. Finally, we construct large-scale nanocapsules with pH-responsive subunits using hierarchical assembly of individual DNA origami monomers. Together, these works expand the foundation for the therapeutic application of DNA origami.