A label-free light-scattering method to resolve assembly and disassembly of DNA nanostructures

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A1 Alkuperäisartikkeli tieteellisessä aikakauslehdessä

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2022-12-20

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en

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4800-4809

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Biophysical Journal, Volume 121, issue 24

Abstract

DNA self-assembly, and in particular DNA origami, has evolved into a reliable workhorse for organizing organic and inorganic materials with nanometer precision and with exactly controlled stoichiometry. To ensure the intended performance of a given DNA structure, it is beneficial to determine its folding temperature, which in turn yields the best possible assembly of all DNA strands. Here, we show that temperature-controlled sample holders and standard fluorescence spectrometers or dynamic light-scattering setups in a static light-scattering configuration allow for monitoring the assembly progress in real time. With this robust label-free technique, we determine the folding and melting temperatures of a set of different DNA origami structures without the need for more tedious protocols. In addition, we use the method to follow digestion of DNA structures in the presence of DNase I and find strikingly different resistances toward enzymatic degradation depending on the structural design of the DNA object.

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Funding Information: We thank V. Aseyev, E. Riedle, and H. Derondeau for technical assistance and fruitful discussions. We thank A. Kucinic and C.E. Castro, who kindly provided us with staple mixtures and scaffolds for the four structures shown in Fig. 4. Financial support by the Emil Aaltonen Foundation, the Sigrid Jusélius Foundation, the Jane and Aatos Erkko Foundation, and ERC consolidator grant “DNA functional lattices” (project ID: 818635) is gratefully acknowledged. This work was carried out under the Academy of Finland Centers of Excellence Program (2022–2029) in Life-Inspired Hybrid Materials (LIBER), project number (346110). We acknowledge the provision of facilities and technical support by Aalto University Bioeconomy Facilities and OtaNano – Nanomicroscopy Center (Aalto-NMC) and Micronova Nanofabrication Center. The authors declare no competing interests. Publisher Copyright: © 2022 Biophysical Society

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Citation

Ijäs, H, Liedl, T, Linko, V & Posnjak, G 2022, ' A label-free light-scattering method to resolve assembly and disassembly of DNA nanostructures ', Biophysical Journal, vol. 121, no. 24, pp. 4800-4809 . https://doi.org/10.1016/j.bpj.2022.10.036