Abstract
ABSTRACTDNA double helix structure is stabilized by the base-pairing and the base-stacking interactions. Base-stacking interactions originating from hydrophobic interactions between the nucleobases predominantly contribute to the duplex stability. A comprehensive understanding of dinucleotide base-stacking interactions is lacking owing to the unavailability of sensitive techniques that can measure these weak interactions. Earlier studies attempting to address this question only managed to estimate the base-pair stacking interactions, however, disentangling individual base-stacking interactions was enigmatic. By combining multiplexed DNA-PAINT imaging with designer DNA nanostructures, we experimentally measure the free energy of dinucleotide base-stacking at the single-molecule level. Multiplexed imaging enabled us to extract binding kinetics of an imager strand with and without additional dinucleotide stacking interactions in a single imaging experiment, abolishing any effects of experimental variations. The DNA-PAINT data showed that a single additional dinucleotide base-stacking results in as much as 250-fold stabilization of the imager strand binding. We found that the dinucleotide base-stacking energies vary from -1.18 ± 0.17 kcal/mol to -3.57 ± 0.08 kcal/mol for C|T and A|C base-stackings, respectively. We demonstrate the application of base-stacking energetics in designing DNA-PAINT probes for multiplexed super-resolution imaging. Our results will aid in designing functional DNA nanostructures, DNA and RNA aptamers, and facilitate better predictions of the local DNA structure.
Publisher
Cold Spring Harbor Laboratory
Cited by
4 articles.
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