Development of bis-locked nucleic acid (bisLNA) oligonucleotides for efficient invasion of supercoiled duplex DNA

Pedro M D Moreno, Sylvain Geny, Y Vladimir Pabon, Helen Bergquist, Eman M Zaghloul, Cristina S J Rocha, Iulian I Oprea, Burcu Bestas, Samir El Andaloussi, Per T Jørgensen, Erik B Pedersen, Karin E Lundin, Rula Zain, Jesper Wengel, C I Edvard Smith

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Abstrakt

In spite of the many developments in synthetic oligonucleotide (ON) chemistry and design, invasion into double-stranded DNA (DSI) under physiological salt and pH conditions remains a challenge. In this work, we provide a new ON tool based on locked nucleic acids (LNAs), designed for strand invasion into duplex DNA (DSI). We thus report on the development of a clamp type of LNA ON-bisLNA-with capacity to bind and invade into supercoiled double-stranded DNA. The bisLNA links a triplex-forming, Hoogsteen-binding, targeting arm with a strand-invading Watson-Crick binding arm. Optimization was carried out by varying the number and location of LNA nucleotides and the length of the triplex-forming versus strand-invading arms. Single-strand regions in target duplex DNA were mapped using chemical probing. By combining design and increase in LNA content, it was possible to achieve a 100-fold increase in potency with 30% DSI at 450 nM using a bisLNA to plasmid ratio of only 21:1. Although this first conceptual report does not address the utility of bisLNA for the targeting of DNA in a chromosomal context, it shows bisLNA as a promising candidate for interfering also with cellular genes.
OriginalsprogEngelsk
TidsskriftNucleic Acids Research
Vol/bind41
Udgave nummer5
Sider (fra-til)3257-3273
Antal sider17
ISSN0305-1048
DOI
StatusUdgivet - 2013

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Moreno, P. M. D., Geny, S., Pabon, Y. V., Bergquist, H., Zaghloul, E. M., Rocha, C. S. J., Oprea, I. I., Bestas, B., Andaloussi, S. E., Jørgensen, P. T., Pedersen, E. B., Lundin, K. E., Zain, R., Wengel, J., & Smith, C. I. E. (2013). Development of bis-locked nucleic acid (bisLNA) oligonucleotides for efficient invasion of supercoiled duplex DNA. Nucleic Acids Research, 41(5), 3257-3273. https://doi.org/10.1093/nar/gkt007