Spatiotemporal Control over Polynucleotide Brush Growth on DNA Origami Nanostructures

DNA nanotechnology provides an approach to create precise, tunable, and biocompatible nanostructures for biomedical applications. However, the stability of these structures is severely compromised in biological milieu due to their fast degradation by nucleases. Recently, we showed how enzymatic polymerization could be harnessed to grow polynucleotide brushes of tunable length and location on the surface of DNA origami nanostructures, which greatly enhances their nuclease stability. Here, we report on strategies that allow for both spatial and temporal control over polymerization through activatable initiation, cleavage, and regeneration of polynucleotide brushes using restriction enzymes. The ability to site-specifically decorate DNA origami nanostructures with polynucleotide brushes in a spatiotemporally controlled way provides access to smart functionalized DNA architectures with potential applications in drug delivery and supramolecular assembly. Novel strategies allow for both spatial and temporal control of polynucleotide brush modification on DNA origami through activation and regeneration of initiation sites using restriction enzymes in combination with TdT-catalyzed enzymatic polymerization. Spatiotemporal control of brush growth provides access to smart functionalized DNA architectures with potential applications ranging from drug delivery to supracolloidal assembly.+image

Automated summary

DNA nanotechnology allows for the creation of precise and tunable nanostructures for biomedical applications. However, the stability of these structures is compromised in biological environments due to degradation by nucleases. This study demonstrates a strategy to enhance the stability of the structures by utilizing enzymatic polymerization to grow polynucleotide brushes on DNA nanostructures. The ability to control the spatial and temporal growth of these brushes offers potential applications in drug delivery and supramolecular assembly.