Vesicle-Templated Self-Assembly of Programmable Freestanding Multi-μm DNA Shells

In the quest to create increasingly complex synthetic cell-mimicking systems, diverse DNA nanostructures have been developed to coat, permeabilize, sculpt, or otherwise functionalize lipid vesicles or used as scaffolds to direct vesicle growth. Here, we introduce a simple, broadly applicable method to realize freestanding, membrane-mimicking DNA shells: DNA shells are first assembled on giant unilamellar vesicles and then liberated by surfactant-mediated liposome removal, retaining the geometry of their membrane templates. We demonstrate this approach using two distinct DNA tecton classes: a complex barrel-shaped DNA origami and a simple 11-oligonucleotide nanostar-inspired motif. The site-specific addressability of DNA origami structures enables the rational design of binding interfaces, as demonstrated by the controlled formation of multilayer shells. The success of both strategies underscores the feasibility of using different DNA architectures to create tunable, DNA-only shell-like compartments spanning the size range of eukaryotic cells, thereby offering a fundamentally new type of compartmentalization for bottom-up synthetic biology.