title: Ternary complex of plasmid DNA with protamine and γ-polyglutamic acid for biocompatible gene delivery system.
abstract: The purpose of the present study was to investigate the usefulness of the ternary complex with protamine and γ-polyglutamic acid (γ-PGA), which are biodegradable materials for foods and medical products, as a safe gene delivery vector. We formed cationic binary complexes (plasmid DNA (pDNA)/protamine complexes) with high transfection efficiency. The binary complex showed slight toxicity probably related to its total cationic charge. Then, we formed ternary complexes (pDNA/protamine/γ-PGA complexes) by addition of anionic polymer, γ-PGA, and they showed no cytotoxicity. The transfection efficiency of the pDNA/protamine/γ-PGA complexes was as high as that of the pDNA/protamine complexes, although their zeta potentials were different. Inhibition study of the gene expressions in B16-F10 cells suggested that pDNA/protamine complexes were taken up by caveolae-mediated endocytosis and macropinocytosis. On the other hand, pDNA/protamine/γ-PGA complexes were taken up by clathrin-mediated endocytosis and macropinocytosis. Thus, we succeeded in developing the ternary complex as a safe gene delivery vector with biocompatible materials.
authors.affiliation: Department of Hospital Pharmacy, Nagasaki University Hospital.
authors.forename: Kosuke
authors.initials: K
authors.lastname: Kanda
authors.forename: Yukinobu
authors.initials: Y
authors.lastname: Kodama
authors.forename: Tomoaki
authors.initials: T
authors.lastname: Kurosaki
authors.forename: Masanobu
authors.initials: M
authors.lastname: Imamura
authors.forename: Hiroo
authors.initials: H
authors.lastname: Nakagawa
authors.forename: Takahiro
authors.initials: T
authors.lastname: Muro
authors.forename: Norihide
authors.initials: N
authors.lastname: Higuchi
authors.forename: Tadahiro
authors.initials: T
authors.lastname: Nakamura
authors.forename: Takashi
authors.initials: T
authors.lastname: Kitahara
authors.forename: Masayuki
authors.initials: M
authors.lastname: Honda
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Sasaki
mesh_terms: D000818:Animals; D001672:Biocompatible Materials; D045744:Cell Line, Tumor; D002470:Cell Survival; D004247:DNA / Q000008:administration & dosage* / Q000737:chemistry; D018014:Gene Transfer Techniques; D018773:Genes, erbB-1; D051379:Mice; D010957:Plasmids; D011099:Polyglutamic Acid / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000737:chemistry; D011479:Protamines / Q000008:administration & dosage* / Q000737:chemistry
chemical_list: D001672:Biocompatible Materials; D011479:Protamines; C511775:poly(gamma-glutamic acid); D011099:Polyglutamic Acid; D004247:DNA