Protein-Assisted Room-Temperature Assembly of Rigid, Immobile Holliday Junctions and Hierarchical DNA Nanostructures

S. Ramakrishnan, S. Subramaniam, C. Kielar, G. Grundmeier, A.F. Stewart, A. Keller, Molecules 25 (2020) 5099.

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Journal Article | Published | English
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<jats:p>Immobile Holliday junctions represent not only the most fundamental building block of structural DNA nanotechnology but are also of tremendous importance for the in vitro investigation of genetic recombination and epigenetics. Here, we present a detailed study on the room-temperature assembly of immobile Holliday junctions with the help of the single-strand annealing protein Redβ. Individual DNA single strands are initially coated with protein monomers and subsequently hybridized to form a rigid blunt-ended four-arm junction. We investigate the efficiency of this approach for different DNA/protein ratios, as well as for different DNA sequence lengths. Furthermore, we also evaluate the potential of Redβ to anneal sticky-end modified Holliday junctions into hierarchical assemblies. We demonstrate the Redβ-mediated annealing of Holliday junction dimers, multimers, and extended networks several microns in size. While these hybrid DNA–protein nanostructures may find applications in the crystallization of DNA–protein complexes, our work shows the great potential of Redβ to aid in the synthesis of functional DNA nanostructures under mild reaction conditions.</jats:p>
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Molecules
Volume
25
Page
5099
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Ramakrishnan S, Subramaniam S, Kielar C, Grundmeier G, Stewart AF, Keller A. Protein-Assisted Room-Temperature Assembly of Rigid, Immobile Holliday Junctions and Hierarchical DNA Nanostructures. Molecules. 2020;25:5099. doi:10.3390/molecules25215099
Ramakrishnan, S., Subramaniam, S., Kielar, C., Grundmeier, G., Stewart, A. F., & Keller, A. (2020). Protein-Assisted Room-Temperature Assembly of Rigid, Immobile Holliday Junctions and Hierarchical DNA Nanostructures. Molecules, 25, 5099. https://doi.org/10.3390/molecules25215099
@article{Ramakrishnan_Subramaniam_Kielar_Grundmeier_Stewart_Keller_2020, title={Protein-Assisted Room-Temperature Assembly of Rigid, Immobile Holliday Junctions and Hierarchical DNA Nanostructures}, volume={25}, DOI={10.3390/molecules25215099}, journal={Molecules}, author={Ramakrishnan, Saminathan and Subramaniam, Sivaraman and Kielar, Charlotte and Grundmeier, Guido and Stewart, A. Francis and Keller, Adrian}, year={2020}, pages={5099} }
Ramakrishnan, Saminathan, Sivaraman Subramaniam, Charlotte Kielar, Guido Grundmeier, A. Francis Stewart, and Adrian Keller. “Protein-Assisted Room-Temperature Assembly of Rigid, Immobile Holliday Junctions and Hierarchical DNA Nanostructures.” Molecules 25 (2020): 5099. https://doi.org/10.3390/molecules25215099.
S. Ramakrishnan, S. Subramaniam, C. Kielar, G. Grundmeier, A. F. Stewart, and A. Keller, “Protein-Assisted Room-Temperature Assembly of Rigid, Immobile Holliday Junctions and Hierarchical DNA Nanostructures,” Molecules, vol. 25, p. 5099, 2020.
Ramakrishnan, Saminathan, et al. “Protein-Assisted Room-Temperature Assembly of Rigid, Immobile Holliday Junctions and Hierarchical DNA Nanostructures.” Molecules, vol. 25, 2020, p. 5099, doi:10.3390/molecules25215099.

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