Development of bis-locked nucleic acid (bisLNA) oligonucleotides for efficient invasion of supercoiled duplex DNA
| Autor: | Y. Vladimir Pabon, Burcu Bestas, Sylvain Geny, Samir El Andaloussi, Rula Zain, C. I. Edvard Smith, Per T. Jørgensen, Jesper Wengel, Eman M. Zaghloul, Cristina S. J. Rocha, Helen Bergquist, Iulian I. Oprea, Erik B. Pedersen, Pedro Moreno, Karin E. Lundin |
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| Rok vydání: | 2013 |
| Předmět: |
Base pair
Molecular Sequence Data Oligonucleotides Context (language use) Buffers Biology 010402 general chemistry 01 natural sciences 03 medical and health sciences chemistry.chemical_compound Plasmid Genetics Transition Temperature Strand invasion DNA Cleavage Locked nucleic acid Base Pairing Molecular Biology 030304 developmental biology 0303 health sciences Binding Sites Base Sequence DNA Superhelical Oligonucleotide DNA DNA Restriction Enzymes Hydrogen-Ion Concentration 0104 chemical sciences Biochemistry chemistry Nucleic acid Plasmids |
| Zdroj: | 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, vol. 41, no. 5, pp. 3257-3273 . https://doi.org/10.1093/nar/gkt007 Nucleic Acids Research Nucleic Acids Research; Vol 41 |
| ISSN: | 1362-4962 0305-1048 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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