Single molecule DNA origami nanoarrays with controlled protein orientation

The nanoscale organization of functional (bio)molecules on solid substrates with spatial resolution and single-molecule control—in both position orientation—is great interest for the development next-generation (bio)molecular devices assays. Herein, we report fabrication nanoarrays individual proteins (and dyes) via selective DNA origami nanopatterned surfaces controlled protein orientation. Nanoapertures in metal-coated glass were patterned using focused ion beam lithography; 88% nanoapertures allowed immobilization functionalized structures. Photobleaching experiments dye-functionalized nanostructures indicated that 85% contain a single unit, only 3% exhibiting double occupancy. Using reprogrammed genetic code to engineer into new chemistry allow residue-specific linkage an addressable ssDNA assembled orientation-controlled structures; these then organized arrays exhibited molecule traces. This strategy is general applicability investigation biomolecular events defined configurations orientational control (bio)molecule interest.