Interfacial Packing of DNA Nanostars Regulates Dynamics on Synthetic Cell Membranes

Abstract

DNA nanostructures are emerging as programmable components for engineering synthetic cell membranes, yet how their collective packing and deformability regulate molecular dynamics at membrane interfaces remains poorly understood. Here, we investigate the packing and mobility of DNA nanostars with tunable stiffness on lipid-coated droplets. The negatively charged nanostars spontaneously adsorb onto cationic membranes, and their interfacial packing is changed by the bulk DNA concentration and droplet size, which together determine the number of encapsulated nanostars. Combining fluorescence recovery after photobleaching experiments with coarse-grained simulations, we reveal distinct packing-dynamics relationships for rigid and soft nanostars. For rigid nanostars, diffusion first decreases gradually and then drops sharply with increasing interfacial packing, approaching a dynamically arrested state consistent with jamming-like behavior. In contrast, at equivalent experimental conditions, soft nanostars systematically reach lower interfacial packing fractions and show a weaker decrease in apparent mobility. This behavior is consistent with their weaker membrane affinity, which facilitates adsorption-desorption with the bulk. When this exchange is suppressed in simulations, crowding reduces the lateral diffusion of both nanostar types, but produces distinct dense configurations: soft nanostars become strongly deformed, whereas rigid nanostars largely retain their shape and form interlocked, gear-like arrangements. Nanostar adsorption also impedes lipid diffusion, while differences in membrane affinity result in distinct lipid mobile fractions. These findings reveal how the interplay between nanostar packing and deformability regulates molecular transport at membrane interfaces, providing a physical design principle for tuning lateral fluidity and crowding in artificial cells.