Scaffolded DNA origami permits the programmable building of nanoscale buildings by means of the hybridization of an extended single-stranded scaffold with tons of of quick staple strands. Nonetheless, the reliance on quite a few artificial oligonucleotides stays a key barrier to scalable and cost-effective manufacturing of DNA nanostructures. On this examine, we introduce a long-staple design technique that extends the size of particular person staple strands to 100–200 nucleotides (nt), thereby lowering the whole variety of strands required whereas sustaining meeting effectivity and structural constancy. We reveal that this method is broadly appropriate with quite a lot of origami architectures, together with each manually designed lattice-based buildings and algorithmically generated wireframe geometries, with out requiring adjustments to well-established design workflows. Utilizing consultant 2D and 3D buildings, we present that lengthy staples can assemble effectively below the identical thermal annealing circumstances and Mg2+ concentrations as quick staples, yielding last buildings with comparable morphology. To additional assist organic manufacturing of staple strands, we generated lengthy staples by way of rolling circle amplification (RCA) utilizing custom-designed round templates, every encoding a selected lengthy staple sequence. This modular design permits for versatile and selective synthesis of desired staples, both individually or in pooled codecs. These RCA-derived staples had been efficiently utilized in construction meeting, confirming the feasibility of enzyme-based synthesis for long-staple designs. This modular and adaptable technique provides a sensible route towards scalable fabrication of purposeful DNA nanostructures throughout numerous design frameworks.
