The effect of base pair mismatch on DNA strand displacement

TL;DR

This study investigates how a single base pair mismatch affects DNA strand displacement kinetics, revealing that mismatches stall branch migration and alter displacement pathways, which can inform DNA nanotechnology design.

Contribution

The paper introduces a quantitative model combining branch migration and dissociation, explaining the effects of mismatches on DNA displacement kinetics.

Findings

Mismatches significantly slow down displacement rates.

Displacement occurs via direct dissociation when branch migration stalls.

The proposed model accurately predicts the impact of mismatches.

Abstract

DNA strand displacement is a key reaction in DNA homologous recombination and DNA mismatch repair and is also heavily utilized in DNA-based computation and locomotion. Despite its ubiquity in science and engineering, sequence-dependent effects of displacement kinetics have not been extensively characterized. Here, we measured toehold-mediated strand displacement kinetics using single-molecule fluorescence in the presence of a single base pair mismatch. The apparent displacement rate varied significantly when the mismatch was introduced in the invading DNA strand. The rate generally decreased as the mismatch in the invader was encountered earlier in displacement. Our data indicate that a single base pair mismatch in the invader stalls branch migration, and displacement occurs via direct dissociation of the destabilized incumbent strand from the substrate strand. We combined both branch migration and direct dissociation into a model, which we term, the concurrent displacement model, and used the first passage time approach to quantitatively explain the salient features of the observed relationship. We also introduce the concept of splitting probabilities to justify that the concurrent model can be simplified into a three-step sequential model in the presence of an invader mismatch. We expect our model to become a powerful tool to design DNA-based reaction schemes with broad functionality.