Vesicle-templated self-assembly of programmable freestanding multi-µm DNA shells

In the quest to create increasingly complex synthetic cell-mimicking systems, a wide range of DNA nanostructures have been developed to coat, permeabilize, sculpt, or otherwise functionalize lipid vesicles. In a complementary strategy, DNA architectures have been used as scaffolds to direct the growth of lipid membrane vesicles. Here we introduce a simple and broadly applicable method to realize freestanding, membrane-mimicking DNA shells: DNA shells are first assembled on the outer surface of giant unilamellar vesicles and then liberated by surfactant-mediated liposome removal. The resulting structures faithfully retain the geometry of their membrane template. We demonstrate the approach with two distinct classes of DNA tectons: a complex barrel-shaped DNA origami with programmable inter-subunit interactions, and a simple nanostar-inspired motif composed of only eleven oligonucleotides. The site-specific addressability of the former enable the rational design of binding interfaces, as demonstrated by controlled multilayer formation. The success of both strategies underscores the generality of our approach and the feasibility of creating shell-like compartments from different DNA architectures. This method enables the construction of tunable, DNA-only containers spanning the size range of eukaryotic cells, offering a fundamentally new type of compartmentalization for bottom-up synthetic biology.