title: Ternary complexes of pDNA, polyethylenimine, and gamma-polyglutamic acid for gene delivery systems.
abstract: We discovered a vector coated by gamma-polyglutamic acid (gamma-PGA) for effective and safe gene delivery. In order to develop a useful non-viral vector, we prepared several ternary complexes constructed with pDNA, polyethylenimine (PEI), and various polyanions, such as polyadenylic acid, polyinosinic-polycytidylic acid, alpha-polyaspartic acid, alpha-polyglutamic acid, and gamma-PGA. The pDNA/PEI complex had a strong cationic surface charge and showed extremely high transgene efficiency although it agglutinated with erythrocytes and had extremely high cytotoxicity. Those polyanions changed the positive zeta-potential of pDNA/PEI complex to negative although they did not affect the size. They had no agglutination activities and lower cytotoxicities but most of the ternary complexes did not show any uptake and gene expression; however, the pDNA/PEI/gamma-PGA complex showed high uptake and gene expression. Most of the pDNA/PEI/gamma-PGA complexes were located in the cytoplasm without dissociation and a few complexes were observed in the nuclei. Hypothermia and the addition of gamma-PGA significantly inhibited the uptake of pDNA/PEI/gamma-PGA by the cells, although l-glutamic acid had no effect. These results strongly indicate that the pDNA/PEI/gamma-PGA complex was taken up by gamma-PGA-specific receptor-mediated energy-dependent process. Thus, the pDNA/PEI/gamma-PGA complex is useful as a gene delivery system with high transfection efficiency and low toxicity.
authors.affiliation: Department of Hospital Pharmacy, Nagasaki University Hospital of Medicine and Dentistry, 1-7-1 Sakamoto, Nagasaki 852-8501, Japan.
authors.forename: Tomoaki
authors.initials: T
authors.lastname: Kurosaki
authors.forename: Takashi
authors.initials: T
authors.lastname: Kitahara
authors.forename: Shintaro
authors.initials: S
authors.lastname: Fumoto
authors.forename: Koyo
authors.initials: K
authors.lastname: Nishida
authors.forename: Junzo
authors.initials: J
authors.lastname: Nakamura
authors.forename: Takuro
authors.initials: T
authors.lastname: Niidome
authors.forename: Yukinobu
authors.initials: Y
authors.lastname: Kodama
authors.forename: Hiroo
authors.initials: H
authors.lastname: Nakagawa
authors.forename: Hideto
authors.initials: H
authors.lastname: To
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Sasaki
mesh_terms: D000818:Animals; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D004247:DNA / Q000737:chemistry* / Q000235:genetics; D004912:Erythrocytes / Q000378:metabolism; D018014:Gene Transfer Techniques; D042541:Intracellular Space / Q000378:metabolism; D051379:Mice; D055668:Photochemical Processes; D010957:Plasmids / Q000737:chemistry* / Q000235:genetics; D011094:Polyethyleneimine / Q000737:chemistry* / Q000633:toxicity; D011099:Polyglutamic Acid / Q000031:analogs & derivatives* / Q000737:chemistry / Q000633:toxicity
chemical_list: C511775:poly(gamma-glutamic acid); D011099:Polyglutamic Acid; D011094:Polyethyleneimine; D004247:DNA