Virus-Inspired Membrane Encapsulation of DNA Nanostructures To Achieve In Vivo Stability

Abstract

DNA nanotechnology enables engineering of molecular-scale devices with exquisite control over geometry and site-specific functionalization. This capability promises compelling advantages in advancing nanomedicine; nevertheless, instability in biological environments and innate immune activation remain as obstacles for in vivo application. Natural particle systems (i.e., viruses) have evolved mechanisms to maintain structural integrity and avoid immune recognition during infection, including encapsulation of their genome and protein capsid shell in a lipid envelope. Here we introduce virus-inspired enveloped DNA nanostructures as a design strategy for biomedical applications. Achieving a high yield of tightly wrapped unilamellar nanostructures, mimicking the morphology of enveloped virus particles, required precise control over the density of attached lipid conjugates and was achieved at 1 per ∼180 nm2. Envelopment of DNA nanostructures in PEGylated lipid bilayers conferred protection against nuclease digestion. Immune activation was decreased 2 orders of magnitude below controls, and pharmacokinetic bioavailability improved by a factor of 17. By establishing a design strategy suitable for biomedical applications, we have provided a platform for the engineering of sophisticated, translation-ready DNA nanodevices.

Synthetic biology devices for in vitro and in vivo diagnostics

“…Although there is great potential for such tools, the in vivo susceptibility of DNA-based nanorobots to degradation must be addressed before use in humans; but, encouragingly, nanorobots have been successfully demonstrated in live insects (86). Solutions to resolve degradation issues have been drawn from the architecture of membrane-encapsulated viruses, with lipid bilayer-coated DNA nanostructures displaying dramatically increased in vivo half-life (87). Nanorobots could perhaps be conjugated to flagellated bacteria and "rafted" to targeted tissues via bacterial self-propulsion, similar to a recent study in which the natural tumor-homing ability of Salmonella typhimuria was exploited for the delivery of attached, fluorescently labeled microbeads (88).…”

Supramolecular Wireframe DNA Polyhedra: Assembly and Applications

“…In the case of in vivo applications, some fundamental issues need to be addressed for the polyhedral DNA structures: (1) how to build a more robust DNA polyhedron with increased in vivo circulation time; (2) how to target a specific tumor cell and a subcellular region with a DNA polyhedron, and (3) how to improve cellular uptake efficiency and facilitate lysosomal escape. Some recent literatures [76,78] have provided clues in response to these questions, while more systematic explorations are still needed. Along with the continued developments of structural DNA nanotechnology in biomedicines and nanomaterials science, we can expect some more realistic applications of this new research discipline in the foreseeable future.…”

Beyond the smiley face: applications of structural DNA nanotechnology

“…Inspired by a virus, and a specific application designed for biomedical application use, the NanoOctahedron structure was developed wherein a DNA nanostructure is enveloped in a lipid bilayer [17] . The nanostructure was constructed using traditional 3D DNA origami techniques, involving self-assembly of scaffold DNA and staple strands.…”

“…The lipid-bilayer exhibited less degradation and immune response activation in vitro and in vivo as compared to non-enveloped NanoOctahedrons. Therefore, this nanostructure may prove useful for biomedical applications such as tumor detection [17]. …”