title: In vivo gene transfection via intravitreal injection of cationic liposome/plasmid DNA complexes in rabbits.
abstract: To optimize the in vivo ocular transfection efficiency of plasmid DNA (pDNA)/cationic liposome complexes, N-[1-(2,3-dioleyloxy)propyl]-N,N,N-trimethylammonium chloride (DOTMA)/dioleoylphosphatidylethanolamine (DOPE) (1:1 molar ratio) liposomes and DOTMA/cholesterol (Chol) (1:1 molar ratio) liposomes were prepared with varying amounts of pDNA. pDNA/cationic liposome complexes were intravitreally injected (100 microL) in rabbits, and luciferase activity in the cornea, aqueous humor, iris-ciliary body, lens, vitreous body, and retina was measured. Transfection efficiency of pDNA alone did not change with pDNA ranging from 40 to 85 mg. In contrast, transfection efficiency of pDNA complexed with DOTMA/Chol liposomes significantly increased with the amount of pDNA ranging from 40 to 85 microg (P < 0.05). pDNA complexed with DOTMA/DOPE liposomes could not be prepared with pDNA greater than 60 microg. Among these experiments, pDNA (85 microg) complexed with DOTMA/Chol liposomes (pDNA:cationic liposome charge ratio (- : +) = 1.0:2.0) showed the highest transfection efficiency in the ocular tissue and its transfection-mediated luciferase activity peaked at 3 days. Among the ocular tissues, the highest gene expression was observed in the aqueous humor.
authors.affiliation: Graduate School of Biomedical Sciences, Nagasaki University, 1-14 Bunkyo-machi, 852-8521, Japan.
authors.forename: Shigeru
authors.initials: S
authors.lastname: Kawakami
authors.forename: Ayaka
authors.initials: A
authors.lastname: Harada
authors.forename: Koji
authors.initials: K
authors.lastname: Sakanaka
authors.forename: Koyo
authors.initials: K
authors.lastname: Nishida
authors.forename: Junzo
authors.initials: J
authors.lastname: Nakamura
authors.forename: Toshiyuki
authors.initials: T
authors.lastname: Sakaeda
authors.forename: Nobuhiro
authors.initials: N
authors.lastname: Ichikawa
authors.forename: Mikiro
authors.initials: M
authors.lastname: Nakashima
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Sasaki
mesh_terms: D000818:Animals; D002412:Cations; D002784:Cholesterol / Q000737:chemistry; D004247:DNA / Q000008:administration & dosage* / Q000737:chemistry; D008081:Liposomes; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics; D008297:Male; D010714:Phosphatidylethanolamines / Q000737:chemistry; D010957:Plasmids; D000644:Quaternary Ammonium Compounds / Q000737:chemistry; D011817:Rabbits; D014162:Transfection / Q000379:methods*; D014822:Vitreous Body / Q000378:metabolism*
chemical_list: C410262:1,2-dioleoyl-glycero-3-phosphatidyl ethanolamine; D002412:Cations; D008081:Liposomes; D010714:Phosphatidylethanolamines; D000644:Quaternary Ammonium Compounds; C054005:N-(1-(2,3-dioleyloxy)propyl)-N,N,N-trimethylammonium; D004247:DNA; D002784:Cholesterol; D008156:Luciferases