Kinetics and Thermodynamics of Watson–Crick Base Pairing Driven DNA Origami Dimerization
2016
Zenk, John | Tuntivate, Chanon | Schulman, Rebecca
We investigate the kinetics and thermodynamics of DNA origami dimerization using flat rectangle origami components and different architectures of Watson–Crick complementary single-stranded DNA (“sticky end”) linking strategies. We systematically vary the number of linkers, the length of the sticky ends on the linker, and linker architecture and measure the corresponding yields as well as forward and reverse reaction rate constants through fluorescence quenching assays. Yields were further verified using atomic force microscopy. We calculate values of H° and ΔS° for various interface designs and find nonlinear van’t Hoff behavior, best described by two linear equations, suggesting distinct regimes of dimerization between those with and those without well-formed interfaces. We find that self-assembly reactions can be tuned by manipulating the interface architecture without suffering a loss in yield, even when yield is high, ∼75–80%. We show that the second-order forward reaction rate constant (kₒₙ) depends on both linker architecture and number of linkers used, with typical values on the order of 10⁵–10⁶ (M·s)⁻¹, values that are similar to those of bimolecular association of small, complementary DNA strands. The kₒₙ values are generally non-Arrhenius, tending to increase with decreasing temperature. Finally, we use kinetic and thermodynamic information about the optimal linking architecture to extend the system to an infinite, two-component repeating lattice system and show that we can form micron-sized lattices, with well-formed structures up to 8 μm².
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