title: Deposition gene transfection using bioconjugates of DNA and thermoresponsive cationic homopolymer.
abstract: A poly(N,N-dimethylaminoethylmethacrylate) (PDMAEMA) homopolymer with both thermoresponsive and cationic characteristics was applied to a vector for use in deposition transfection. PDMAEMA with a molecular weight of 2.5 × 10(5) g mol(-1) was synthesized by photoinduced radical polymerization. Polyplexes approximately 750 nm in size were formed by mixing PDMAEMA with luciferase-encoding plasmid DNA. The polyplexes had a lower critical solution temperature (LCST) of approximately 30 °C. In addition, they exhibited excellent adsorption and durability on a polystyrene surface, as confirmed by a surface chemical compositional analysis. When HeLa cells and primary cells were cultured on a substrate coated with the polyplexes, high transgene expression and cell viability of more than 90% were obtained at low charge ratios (PDMAEMA/plasmid DNA ratio) ranging from 2 to 8. In addition, transgene expression was sustained for over 2 weeks post-transfection whereas decreased expression was observed 5 days post-transfection when the conventional solution-mediated transfection method was used. Thus, high and sustained transgene expression as well as high cell viability can be realized by using small amounts of PDMAEMA as a deposition transfection material.
authors.affiliation: Division of Medical Engineering and Materials, National Cerebral and Cardiovascular Center Research Institute, Japan.
authors.forename: Ryosuke
authors.initials: R
authors.lastname: Iwai
authors.forename: Shota
authors.initials: S
authors.lastname: Kusakabe
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Nemoto
authors.forename: Yasuhide
authors.initials: Y
authors.lastname: Nakayama
mesh_terms: D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D004247:DNA / Q000737:chemistry* / Q000235:genetics*; D004285:Dogs; D004337:Drug Carriers / Q000737:chemistry* / Q000633:toxicity; D006367:HeLa Cells; D006801:Humans; D008689:Methacrylates / Q000737:chemistry* / Q000633:toxicity; D009757:Nylons / Q000737:chemistry* / Q000633:toxicity; D013501:Surface-Active Agents / Q000737:chemistry* / Q000633:toxicity; D013696:Temperature; D014162:Transfection / Q000379:methods*
chemical_list: D004337:Drug Carriers; D008689:Methacrylates; D009757:Nylons; D013501:Surface-Active Agents; C407037:poly(2-(dimethylamino)ethyl methacrylate); D004247:DNA