Robustness of localized DNA strand displacement cascades

Colocalization can strongly alter the kinetics and efficiency of chemical processes. For instance, in DNA-templated synthesis unfavorable reactions are sped up by placing reactants into close proximity onto a DNA scaffold. In biochemistry, clustering of enzymes has been demonstrated to enhance the reaction flux through some enzymatic cascades. Here we investigate the effect of colocalization on the performance of DNA strand displacement (DSD) reactions, an important class of reactions utilized in dynamic DNA nanotechnology. We study colocalization by immobilizing a two-stage DSD reaction cascade comprised of a “sender” and a “receiver” gate onto a DNA origami platform. The addition of a DNA (or RNA) input strand displaces a signal strand from the sender gate, which can then transfer to the receiver gate. The performance of the cascade is found to vary strongly with the distance between the gates. A cascade with an intermediate gate distance of ≈20 nm exhibits faster kinetics than those with larger distances, whereas a cascade with smaller distance is corrupted by excessive intraorigami leak reactions. The 20 nm cascade is found to be considerably more robust with respect to a competing reaction, and implementation of multiple receiver gates further increases this robustness. Our results indicate that for the 20 nm distance a fraction of signal strands is transferred locally to a receiver gate on the same platform, probably involving direct physical contact between the gates. The performance of the cascade is consistent with a simple model that takes “local” and “global” transfer processes into account.