화학공학소재연구정보센터
Journal of Physical Chemistry B, Vol.121, No.16, 4015-4022, 2017
Selective and Robust Stabilization of Triplex DNA Structures Using Cationic Comb-type Copolymers
DNA sequences capable of forming triplexes induce DNA,double strand breaks that have attracted attention in genome editing technologies (e.g., CRISPR/Cas9 system, TALEN, and ZEN). Therefore, novel fUnctional tools that stabilize triplex DNA Structures' must be further investigated to spark renewed interest. In this study, we investigated the unique, character of cationic col-Ph-type 'copolymers for the selective stabilization of triplex DNA. The melting temperature (Tm) of triplex DNA increased from 24.5 to 73.0 degrees C (ATm = 48.5 degrees C) by the addition of poly(allylamine)-graft-dextran (PAA-g-Dex) under physiological.. conditions (at pH 7.0), while PAA-g-Dex did not stabilize but rather destabilized the DNA duplex. On the other hand, poly(Oysine)-graft-dextran (PLL-g-Dex) stabilized both the duplex and triplex structures at pH 7.0. Thermodynamic parameters evaluated by isothermal titration calorimetry (ITC) revealed that the binding constant (K-a) for the intermolecular triplex formation in the presence of PAA-g-Dex was 1.1 X 10(9) M-1 at 25 degrees C Which is more than 10 times larger than that in the4Dresence of PLL-g-Dex (8.6 x 10(7) M-1). Stabilizing activity and selectivity of cationic copolymers toward DNA assemblies were-successfully controlled by selecting appropriate"backbone structures of the copolymer. Various functional molecules that stabilize DNA duplexes-have been developed and used in biological research. However, there are few cationic polymers that stabilize triplex DNA selectively. This study indicates that PAA-g-Dex has great potential to regulate the biological activities of triplex DNA.