Ice nucleation by DNA origami

Fundamental investigations of ice nucleation, a key process in fields from environmental science to cryobiology, require model systems with chemical and physical structures that are well defined and easily varied. DNA origami is an especially promising model because of the exquisite control that it offers over the physical geometry of the nucleating agent at the nano-scale. Here we compare ice nucleation by solutions of a rectangular DNA origami tile, formed by annealing a 2.6 kbase single-stranded DNA scaffold with ninety shorter 'staple' oligonucleotides, to ice nucleation when these components are mixed at the same concentrations but not annealed. Isothermal measurements show that the molecular conformation has a dramatic effect on the ice nucleating efficiency. For an array of droplets containing annealed, well-folded tiles the freezing rate is constant, whereas for unannealed DNA the freezing rate decreases with time. Despite the freezing rate measured at low temperature being higher for the annealed DNA origami samples than for a significant proportion of the unannealed ones, in slow temperature-ramp measurements the latter generally freeze at higher temperatures. We show that this behaviour is consistent with the formation of small numbers of highly efficient nucleating agents in the unannealed samples, likely through molecular aggregation.