2'-O,4'-C-ethylene bridged nucleic acid modification enhances pyrimidine motif triplex-forming ability under physiological condition

• Published in: Journal of biochemistry (Tokyo) 2012-07, Vol.152 (1), p.17-26
• Main Authors: Torigoe, Hidetaka, Sato, Norihiro, Nagasawa, Nobuyuki
• Format: Article
• Language: English
• Subjects: Base Sequence; Binding Sites; Circular Dichroism; DNA - chemistry; Ethylenes - chemistry; Hydrogen-Ion Concentration; Kinetics; Molecular Sequence Data; Nucleic Acid Conformation; Oligonucleotides - chemistry; Pyrimidines - chemistry; Thermodynamics
• ISSN: 0021-924X; 1756-2651
• DOI: 10.1093/jb/mvs049

Since pyrimidine motif triplex DNA is unstable at physiological neutral pH, triplex stabilization at physiological neutral pH is important for improvement of its potential to be applied to various methods in vivo, such as repression of gene expression, mapping of genomic DNA and gene-targeted mutagenesis. For this purpose, we studied the thermodynamic and kinetic effects of a chemical modification, 2'-O,4'-C-ethylene bridged nucleic acid (ENA) modification of triplex-forming oligonucleotide (TFO), on pyrimidine motif triplex formation at physiological neutral pH. Thermodynamic investigations indicated that the modification achieved more than 10-fold increase in the binding constant of the triplex formation. The increased number of the modification in TFO enhanced the increased magnitude of the binding constant. On the basis of the obtained thermodynamic parameters, we suggested that the remarkably increased binding constant by the modification may result from the increased stiffness of TFO in the unbound state. Kinetic studies showed that the considerably decreased dissociation rate constant resulted in the observed increased binding constant by the modification. We conclude that ENA modification of TFO could be a useful chemical modification to promote the triplex formation under physiological neutral condition, and may advance various triplex formation-based methods in vivo.