Fig. 1: Design and assembly of a DNA-based Nano-winch.

A Schematic illustration of the assembly strategy of the DNA-based Nano-winch. Double-helical DNA domains are represented by cylinders and are packed on a honeycomb lattice. The Nano-winch comprises two origami in a 1:2 trimer, a central Piston-cylinder and two Landing Legs. The central piston-cylinder core origami has eight strands to anneal to the inner face of the Landing Leg origami (i). To prevent toppling, two ~30 nm six-helix bundles project at 90° from the origami landing legs and 45° away from each other to lay parallel to a surface and maximize area coverage to retain an upright position. They are reinforced with a dsDNA strut. Single-stranded DNA connectors link the top and bottom of the cylinder to the backstop and the piston tip, respectively (ii). These single-stranded DNA connectors loops act as entropic springs with stiffness kDNA and exert defined force which is mechanically translated through the rigid origami to the tip of the piston coupled to a molecular target with stiffness kprotein. The length of these connectors can be adjusted by storing the excess scaffold in reservoir loops on the backstop (iii). The tip of the piston positions up to three ligand moieties targeting specific cell surface receptors (iv). Extension of the Piston results in equivalent and opposing compressive force through the Landing Legs. B 1.0% agarose gel with 11 mM MgCl₂ on which the following samples were electrophoresed: M, 1-kb ladder, Piston-cylinder monomer, Landing Leg monomer, and Piston-cylinder incubated with two-fold molar excess of Landing Legs with a concentration gradient from 11 mM to 35 mM MgCl₂. Schematic representations and reference-free class averages from single-particle TEM micrographs of individual components (particle sets available in Supplementary Figs. 5, 8, and 9). Fully assembled Nano-winch particles were visualized both laterally and from above. C Each landing leg can be modified with eight cholesterol moieties, for a total of 32-modifications. Example TEM images of Nano-winches functionalized with cholesterol moieties adhering to small unilamellar vesicles through the landing legs. Distortions of liposomes is an effect of the process of sample preparation for negative stain TEM. White bars represent 50 nm. All TEM analyses were conducted at least three times for each sample. Source data are provided as a Source Data file.