Prescribing DNA Origami Barrel‐Directed Subtractive Patterning of Nanoparticles for Crystalline Superstructure Assembly

Abstract Long‐range ordered lattices formed by the directed arrangement of colloidal particles hold significant promise for applications such as photonic crystals, plasmonic metamaterials, and semiconductor electronics. Harnessing regioselective interactions through DNA‐mediated assembly is a promising approach to advancing colloidal assembly. Despite efforts to engineer microscale patchy particles using sequence‐specific binding properties of DNA, the control of patch formation on nanoscale isotropic spherical nanoparticles remains challenging. We demonstrate a subtractive patterning strategy using barrel‐shaped DNA origami (DNA barrel) to selectively block surfaces of DNA‐coated gold nanospheres and create regiospecific patches. By designing binding positions and geometric parameters of DNA barrels, we can achieve controlled accessibility to nanosphere surfaces, forming patchy nanoparticles with tunable patch numbers and sizes. This strategy enables the construction of multidimensional superstructures with well‐defined stereo relationships, represented by an unprecedented graphane‐like bilayered superlattice. Furthermore, we developed a geometrical model that accounts for anisotropic particle bonding and steric hindrance, elucidating the relationship between architectural outcomes and the structural parameters of DNA‐barrel‐directed patchy nanoparticles, and enabling reverse engineering designs of potential assembly symmetries. This approach opens new avenues for generating nanoparticle assemblies with distinct symmetries and properties.