title: A novel, bio-reducible gene vector containing arginine and histidine enhances gene transfection and expression of plasmid DNA.
abstract: We have engineered a novel, non-viral, multifunctional gene vector (STR-CH(2)R(4)H(2)C) that contained stearoyl (STR) and a block peptide consisting of Cys (C), His (H), and Arg (R). STR-CH(2)R(4)H(2)C can form a stable nano-complex with plasmid DNA (pDNA) based on electronic interactions and disulfide cross linkages. In this study, we evaluated the efficacy of STR-CH(2)R(4)H(2)C as a gene vector. We first determined the optimal weight ratio for STR-CH(2)R(4)H(2)C/pDNA complexes. The complexes with a weight ratio of 50 showed the highest transfection efficacy. We also examined the transfection efficacy of STR-CH(2)R(4)H(2)C/pDNA complexes with or without serum and compared STR-CH(2)R(4)H(2)C/pDNA transfection efficacy with that of Lipofectamine. Even in the presence of serum, STR-CH(2)R(4)H(2)C showed higher transfection efficacy than did Lipofectamine. In addition, we determined the mechanism of transfection of the STR-CH(2)R(4)H(2)C/pDNA complexes using various cellular uptake inhibitors and evaluated its endosomal escape ability using chloroquine. Macropinocytosis was main cellular uptake pathway of STR-CH(2)R(4)H(2)C/pDNA complexes. Our results suggested that STR-CH(2)R(4)H(2)C is a promising gene delivery system.
authors.affiliation: Laboratory of Pharmaceutics and Drug Delivery, Department of Pharmaceutical Science, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, Hachioji, Tokyo, Japan.
authors.forename: Ko
authors.initials: K
authors.lastname: Tanaka
authors.forename: Takanori
authors.initials: T
authors.lastname: Kanazawa
authors.forename: Takaya
authors.initials: T
authors.lastname: Ogawa
authors.forename: Yumiko
authors.initials: Y
authors.lastname: Suda
authors.forename: Yuuki
authors.initials: Y
authors.lastname: Takashima
authors.forename: Tsunehiko
authors.initials: T
authors.lastname: Fukuda
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Okada
mesh_terms: D000818:Animals; D001120:Arginine / Q000008:administration & dosage / Q000737:chemistry* / Q000235:genetics; D019556:COS Cells; D002522:Chlorocebus aethiops; D018076:DNA, Complementary / Q000008:administration & dosage / Q000737:chemistry* / Q000235:genetics; D005822:Genetic Vectors / Q000008:administration & dosage / Q000737:chemistry* / Q000235:genetics; D006639:Histidine / Q000008:administration & dosage / Q000737:chemistry* / Q000235:genetics; D010957:Plasmids / Q000008:administration & dosage / Q000737:chemistry* / Q000235:genetics; D014162:Transfection / Q000379:methods*; D015854:Up-Regulation / Q000235:genetics*
chemical_list: D018076:DNA, Complementary; D006639:Histidine; D001120:Arginine