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Natural and Modified Oligonucleotide Sequences Show Distinct Strand Displacement Kinetics and These Are Affected Further by Molecular Crowders

1
Laboratory of Experimental and Translational Oncology, Department of Oncology, Microbiology and Immunology, Faculty of Science and Medicine, University of Fribourg, Chemin du Musée 18, PER17, 1700 Fribourg, Switzerland
2
BioPhysics, Adolphe Merkle Institute, University of Fribourg, Chemin des Verdiers 4, PER 18, 1700 Fribourg, Switzerland
*
Author to whom correspondence should be addressed.
Academic Editor: Michael Van Dyke
Biomolecules 2022, 12(9), 1249; https://doi.org/10.3390/biom12091249
Received: 27 July 2022 / Revised: 2 September 2022 / Accepted: 4 September 2022 / Published: 6 September 2022
(This article belongs to the Section Biomacromolecules: Nucleic Acids)
DNA and RNA strand exchange is a process of fundamental importance in biology. Herein, we used a FRET-based assay to investigate, for the first time, the stand exchange kinetics of natural DNA, natural RNA, and locked nucleic acid (LNA)-modified DNA sequences in vitro in PBS in the absence or presence of molecular additives and macromolecular crowders such as diethylene glycol dimethyl ether (deg), polyethylene glycol (peg), and polyvinylpyrrolidone (pvp). The results show that the kinetics of strand exchange mediated by DNA, RNA, and LNA-DNA oligonucleotide sequences are different. Different molecular crowders further affect the strand displacement kinetics, highlighting the complexity of the process of nucleic acid strand exchange as it occurs in vivo. In a peg-containing buffer, the rate constant of displacement was slightly increased for the DNA displacement strand, while it was slightly decreased for the RNA and the LNA-DNA strands compared with displacement in pure PBS. When we used a deg-containing buffer, the rate constants of displacement for all three sequences were drastically increased compared with displacement in PBS. Overall, we show that interactions of the additives with the duplex strands have a significant effect on the strand displacement kinetics and this effect can exceed the one exerted by the chemical nature of the displacement strand itself. View Full-Text
Keywords: nucleic acids; strand displacement; hybridization; FRET nucleic acids; strand displacement; hybridization; FRET
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MDPI and ACS Style

Domljanovic, I.; Ianiro, A.; Rüegg, C.; Mayer, M.; Taskova, M. Natural and Modified Oligonucleotide Sequences Show Distinct Strand Displacement Kinetics and These Are Affected Further by Molecular Crowders. Biomolecules 2022, 12, 1249. https://doi.org/10.3390/biom12091249

AMA Style

Domljanovic I, Ianiro A, Rüegg C, Mayer M, Taskova M. Natural and Modified Oligonucleotide Sequences Show Distinct Strand Displacement Kinetics and These Are Affected Further by Molecular Crowders. Biomolecules. 2022; 12(9):1249. https://doi.org/10.3390/biom12091249

Chicago/Turabian Style

Domljanovic, Ivana, Alessandro Ianiro, Curzio Rüegg, Michael Mayer, and Maria Taskova. 2022. "Natural and Modified Oligonucleotide Sequences Show Distinct Strand Displacement Kinetics and These Are Affected Further by Molecular Crowders" Biomolecules 12, no. 9: 1249. https://doi.org/10.3390/biom12091249

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