Enzyme‐Powered DNA Origami Nanostructures for Enhanced Mucosal Diffusion

Crossing mucosal barriers is a central challenge for oral drug delivery, where nanoparticle design must balance stability with mobility in complex fluids. Here, we demonstrate DNA origami as a programmable platform to investigate these processes. Using Förster resonance energy transfer analysis, we show that DNA nanostructures retain their structural integrity for extended periods in porcine intestinal fluid and mucus, establishing their suitability for biologically relevant environments. Building on this, we used single‐particle tracking to assess enzyme‐powered propulsion within mucus. Both urease and catalase induced enhanced diffusion only when anchored to the DNA origami structure, with propulsion persisting for tens of minutes. Importantly, the spatial organization of enzymes dictated performance: symmetric placement of urease enhanced mobility via uniform local pH gradients, whereas asymmetric catalase placement enabled efficient bubble‐driven propulsion. These results highlight DNA origami as a uniquely versatile tool to dissect structure‐function relationships in mucus transport and provide design principles for next‐generation, enzyme‐powered oral delivery systems.