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

Abstract:

: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.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Moreno PM,Geny S,Pabon YV,Bergquist H,Zaghloul EM,Rocha CS,Oprea II,Bestas B,Andaloussi SE,Jørgensen PT,Pedersen EB,Lundin KE,Zain R,Wengel J,Smith CI

doi

10.1093/nar/gkt007

subject

Has Abstract

pub_date

2013-03-01 00:00:00

pages

3257-73

issue

5

eissn

0305-1048

issn

1362-4962

pii

gkt007

journal_volume

41

pub_type

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