Photo-Control of the Polyplexes Formation between DNA and Photo- Cation Generatable Water-Soluble Polymers

Mariko      Umeda; Mariko      Harada-Shiba; Kingo      Uchida; Yasuhide      Nakayama

doi:10.2174/1567201054367986

Abstract

Photo-cation generatable water-soluble polymers (Mw: approximately 1 x 10 5) were prepared by radical copolymerization of N,N-dimethylacrylamide and vinyl monomer of triphenylmethane leucohydroxide (malachite green), which generate a cation upon ultraviolet light (UV) irradiation at wavelengths of between 290 and 410 nm. The malachite green contents of the copolymers were 3.6 (0.4 mol %), 7.9 (0.7 mol %), and 15.0 (2.7 mol %) per molecule. When an aqueous solution of the photo-cationized copolymers generated by UV irradiation was mixed with a Tris-HCl buffer (pH 7.4) of DNA (pGL3-control plasmid), dynamic light scattering (DLS) measurements showed the formation of polyplexes between the cationic copolymers and anionic DNA by non-specific electrostatic interaction, which was visualized with a confocal laser scanning microscopy (CLMS). Their mean cumulant diameter was about 150 nm with low polydispersity irrespective of the malachite green content in the copolymers. In the copolymer with the lowest malachite green content, almost all of the amount of the polyplexes was maintained by repeated UV irradiation, but the amount gradually decreased in the dark at 37°C due to dissociation of the polyplexes, synchronized with the neutralization of the cation form of malachite green. The photo-cation generatable copolymers designed here can undergo photo-induced formation of the polyplexes with DNA and thermal polyplex dissociation, which may be used as a model for a novel photo-induced gene delivery system into cells.

Keywords: photo-induced polyplex formation, photo-control, photo-cation generatable polymer, malachite green, polyplex dissociation