Bottom-up assembly of functional DNA-based cytoskeletons for synthetic cells

Bottom-up synthetic biology aims at reconstructing a cell from biomolecular constituents. However, the combination of multiple elements and functions remained elusive, which stimulates endeavors to explore entirely synthetic bio-inspired solutions towards engineering life. To this end, DNA nanotechnology represents one of the most promising routes, given the inherent sequence specificity, addressability, and programmability of DNA. Here, we demonstrate functional DNA-based cytoskeletons operating in microfluidic cell-sized compartments and lipid vesicles. The synthetic cytoskeletons consist of DNA tiles self-assembled into filament networks. These filaments can be rationally designed and controlled to imitate features of natural cytoskeletons, including dynamic instability, ATP-triggered polymerization, and vesicle transport in cell-sized confinement. Also, they possess engineerable characteristics, including assembly and disassembly powered by DNA hybridization, light or aptamer-target interactions and autonomous transport of gold nanoparticles. This work underpins DNA nanotechnology as a key player in building synthetic cells.