title: Selective and Robust Stabilization of Triplex DNA Structures Using Cationic Comb-type Copolymers.
abstract: 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 ZFN). 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 comb-type copolymers for the selective stabilization of triplex DNA. The melting temperature (Tm) of triplex DNA increased from 24.5 to 73.0 °C (ΔTm= 48.5 °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(l-lysine)-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 (Ka) for the intermolecular triplex formation in the presence of PAA-g-Dex was 1.1 × 109M-1at 25 °C which is more than 10 times larger than that in the presence of PLL-g-Dex (8.6 × 107M-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.
authors.affiliation: The Hakubi Center for Advanced Research, Kyoto University , Yoshida-ushinomiyacho, Sakyo-ku, Kyoto 606-8501, Japan.|Department of Chemistry, Graduate School of Science, Kyoto University , Kitashirakawa-oiwakecho, Sakyo-ku, Kyoto 606-8502, Japan.
authors.forename: Asako
authors.identifier: 0000-0001-7971-6797
authors.initials: A
authors.lastname: Yamayoshi
authors.affiliation: Faculty of Frontiers of Innovative Research in Science and Technology (FIRST), Konan University , 7-1-20 Minatojima-minamimachi, Chuo-ku, Kobe 650-0047, Japan.
authors.forename: Daisuke
authors.initials: D
authors.lastname: Miyoshi
authors.affiliation: Faculty of Frontiers of Innovative Research in Science and Technology (FIRST), Konan University , 7-1-20 Minatojima-minamimachi, Chuo-ku, Kobe 650-0047, Japan.
authors.forename: Yu-Ki
authors.initials: YK
authors.lastname: Zouzumi
authors.affiliation: Department of Biomolecular Engineering, Graduate School of Science and Technology, Kyoto Institute of Technology , Matsugasaki, Sakyo-ku, Kyoto 606-8585, Japan.
authors.forename: Yohei
authors.initials: Y
authors.lastname: Matsuyama
authors.affiliation: Department of Chemistry, Graduate School of Science, Kyoto University , Kitashirakawa-oiwakecho, Sakyo-ku, Kyoto 606-8502, Japan.|Department of Biomolecular Engineering, Graduate School of Science and Technology, Kyoto Institute of Technology , Matsugasaki, Sakyo-ku, Kyoto 606-8585, Japan.
authors.forename: Jumpei
authors.initials: J
authors.lastname: Ariyoshi
authors.affiliation: Department of Life Science and Technology, Tokyo Institute of Technology , 4259 B-57 Nagatsuta, Midori-ku, Yokohama, Kanagawa 226-8501, Japan.
authors.forename: Naohiko
authors.initials: N
authors.lastname: Shimada
authors.affiliation: Department of Biomolecular Engineering, Graduate School of Science and Technology, Kyoto Institute of Technology , Matsugasaki, Sakyo-ku, Kyoto 606-8585, Japan.
authors.forename: Akira
authors.initials: A
authors.lastname: Murakami
authors.affiliation: Institute of Multidisciplinary Research for Advanced Materials, Tohoku University , 2-1-1, Katahira, Aoba-ku, Sendai 980-8577, Japan.
authors.forename: Takehiko
authors.initials: T
authors.lastname: Wada
authors.affiliation: Department of Life Science and Technology, Tokyo Institute of Technology , 4259 B-57 Nagatsuta, Midori-ku, Yokohama, Kanagawa 226-8501, Japan.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Maruyama
mesh_terms: D004247:DNA / Q000737:chemistry*; D003911:Dextrans / Q000737:chemistry / Q000494:pharmacology*; D005079:Excipients / Q000737:chemistry / Q000494:pharmacology*; D008958:Models, Molecular; D009690:Nucleic Acid Conformation / Q000187:drug effects; D009691:Nucleic Acid Denaturation / Q000187:drug effects; D011073:Polyamines / Q000737:chemistry / Q000494:pharmacology*; D011107:Polylysine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000494:pharmacology; D013816:Thermodynamics
chemical_list: D003911:Dextrans; D005079:Excipients; D011073:Polyamines; C113090:poly(lysine)-graft-dextran; C051802:triplex DNA; D011107:Polylysine; C063994:polyallylamine; D004247:DNA