Modular Assembly of Higher-Order DNA Nanotube Tile Nanostructures Using DNA Annular Scaffolds

Inspired by natural systems, synthetic biology employs diverse biomaterials to construct artificial systems on the basis of the principle of modular assembly. However, the rational construction of predictable, stable, and logically controllable higher-order nanostructures remains challenging. DNA nanotechnology, particularly DNA tile and DNA origami strategies, has enabled advances for the construction of artificial modular systems. Nevertheless, the modular potential of the existing DNA assembly strategies has yet to be fully realized. Thus, an assembly strategy that integrates the merits of DNA tiles and DNA origami is required to overcome current limitations in higher-order structure fabrication. Here, we propose a modular assembly strategy based on programmable DNA nanotube (DNT) tiles as building blocks to construct high-order, modular systems. The DNT tiles ensure stability and integrity through multiple DNA annular scaffolds interconnected with short DNA motifs, achieving an assembly yield of over 99% at different diameters. By rationally tuning scaffold size, monomer stoichiometry, and branched junction motifs, we achieved modular assembly, ranging from 30 homogeneous and heterogeneous polymers, Y-shaped and cross-shaped superstructures, to linear oligomers. Leveraging the addressability and dynamic reconfigurability of the modular components, we further constructed a series of DNA-based Boolean logic gates and circuits, demonstrating rapid disassembly and component reusability. This modular strategy expands the design space of limitedly addressable DNA nanostructures and provides a framework for application-oriented DNA nanotechnologies.