title: Secure and effective gene delivery system of plasmid DNA coated by polynucleotide.
abstract: Polynucleotides are anionic macromolecules which are expected to transfer into the targeted cells through specific uptake mechanisms. So, we developed polynucleotides coating complexes of plasmid DNA (pDNA) and polyethylenimine (PEI) for a secure and efficient gene delivery system and evaluated their usefulness. Polyadenylic acid (polyA), polyuridylic acid (polyU), polycytidylic acid (polyC), and polyguanylic acid (polyG) were examined as the coating materials. pDNA/PEI/polyA, pDNA/PEI/polyU, and pDNA/PEI/polyC complexes formed nanoparticles with a negative surface charge although pDNA/PEI/polyG was aggregated. The pDNA/PEI/polyC complex showed high transgene efficiency in B16-F10 cells although there was little efficiency in pDNA/PEI/polyA and pDNA/PEI/polyU complexes. An inhibition study strongly indicated the specific uptake mechanism of pDNA/PEI/polyC complex. Polynucleotide coating complexes had lower cytotoxicity than pDNA/PEI complex. The pDNA/PEI/polyC complex showed high gene expression selectively in the spleen after intravenous injection into mice. The pDNA/PEI/polyC complex showed no agglutination with erythrocytes and no acute toxicity although these were observed in pDNA/PEI complex. Thus, we developed polynucleotide coating complexes as novel vectors for clinical gene therapy, and the pDNA/PEI/polyC complex as a useful candidate for a gene delivery system.
authors.affiliation: Department of Hospital Pharmacy, Nagasaki University Hospital , Nagasaki , Japan and.
authors.forename: Yukinobu
authors.initials: Y
authors.lastname: Kodama
authors.forename: Chikako
authors.initials: C
authors.lastname: Ohkubo
authors.forename: Tomoaki
authors.initials: T
authors.lastname: Kurosaki
authors.forename: Kanoko
authors.initials: K
authors.lastname: Egashira
authors.forename: Kayoko
authors.initials: K
authors.lastname: Sato
authors.forename: Shintaro
authors.initials: S
authors.lastname: Fumoto
authors.forename: Koyo
authors.initials: K
authors.lastname: Nishida
authors.forename: Norihide
authors.initials: N
authors.lastname: Higuchi
authors.forename: Takashi
authors.initials: T
authors.lastname: Kitahara
authors.forename: Tadahiro
authors.initials: T
authors.lastname: Nakamura
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Sasaki
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D004247:DNA / Q000008:administration & dosage* / Q000737:chemistry*; D004912:Erythrocytes / Q000187:drug effects / Q000378:metabolism; D015870:Gene Expression; D018014:Gene Transfer Techniques / Q000009:adverse effects; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000008:administration & dosage / Q000009:adverse effects / Q000737:chemistry*; D051379:Mice; D053758:Nanoparticles / Q000008:administration & dosage / Q000737:chemistry; D010316:Particle Size; D010957:Plasmids / Q000008:administration & dosage* / Q000737:chemistry; D011094:Polyethyleneimine / Q000008:administration & dosage / Q000737:chemistry; D011119:Polynucleotides / Q000008:administration & dosage / Q000009:adverse effects / Q000737:chemistry*; D013154:Spleen / Q000378:metabolism; D055672:Static Electricity; D013499:Surface Properties
chemical_list: D011119:Polynucleotides; D011094:Polyethyleneimine; D004247:DNA
keywords: Gene delivery; pDNA; polyethylenimine; polynucleotide; ternary complex