title: DNA amplification in neutral liposomes for safe and efficient gene delivery.
abstract: In general, traditional gene carriers contain strong cationic charges to efficiently load anionic genes, but this cationic character also leads to destabilization of plasma membranes and causes severe cytotoxicity. Here, we developed a PCR-based nanofactory as a safe gene delivery system. A few template plasmid DNA can be amplified by PCR inside liposomes about 200 nm in diameter, and the quantity of loaded genes highly increased by more than 8.8-fold. The liposome membrane was composed of neutral lipids free from cationic charges. Consequently, this system is nontoxic, unlike other traditional cationic gene carriers. Intense red fluorescent protein (RFP) expression in CHO-K1 cells showed that the amplified genes could be successfully transfected to cells. Animal experiments with the luciferase gene also showed in vivo gene expression by our system without toxicity. We think that this PCR-based nanofactory system can overcome the toxicity problem that is the critical limitation of current gene delivery to clinical application.
authors.affiliation: Center for Theragnosis, Biomedical Research Institute, Korea, Institute of Science and Technology , 39-1 Hawolgok-dong, Seongbuk-gu, Seoul 136-791, Republic of Korea.
authors.forename: Sangmin
authors.initials: S
authors.lastname: Lee
authors.forename: Heebeom
authors.initials: H
authors.lastname: Koo
authors.forename: Jin Hee
authors.initials: JH
authors.lastname: Na
authors.forename: Kyung Eun
authors.initials: KE
authors.lastname: Lee
authors.forename: Seo Young
authors.initials: SY
authors.lastname: Jeong
authors.forename: Kuiwon
authors.initials: K
authors.lastname: Choi
authors.forename: Sun Hwa
authors.initials: SH
authors.lastname: Kim
authors.forename: Ick Chan
authors.initials: IC
authors.lastname: Kwon
authors.forename: Kwangmeyung
authors.initials: K
authors.lastname: Kim
mesh_terms: D000818:Animals; D000838:Anions; D016466:CHO Cells; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism; D002470:Cell Survival; D006224:Cricetinae; D003412:Cricetulus; D004247:DNA / Q000737:chemistry*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000737:chemistry; D006801:Humans; D018664:Interleukin-12 / Q000378:metabolism; D008055:Lipids / Q000737:chemistry; D008081:Liposomes / Q000737:chemistry*; D008164:Luminescent Proteins / Q000737:chemistry; D008856:Microscopy, Fluorescence; D053758:Nanoparticles / Q000737:chemistry; D036103:Nanotechnology / Q000379:methods*; D009369:Neoplasms / Q000378:metabolism; D016133:Polymerase Chain Reaction; D014162:Transfection; D000097573:Red Fluorescent Protein
chemical_list: D000838:Anions; D004247:DNA; D018664:Interleukin-12; D008055:Lipids; D008081:Liposomes; D008164:Luminescent Proteins; D000097573:Red Fluorescent Protein