title: Versatile nuclear localization signal-based oligopeptide as a gene vector.
abstract: To develop a versatile nuclear-targeted gene vector, nuclear localization signal (NLS) oligopeptides combining cysteine (C), histidine (H), and stearic acid (STR) were investigated in this study. The original SV40 sequence (SV40: Pro-Lys-Lys-Lys-Arg-Lys-Val) was selected as the NLS sequence, and physical characterizations of various NLS-based oligopeptides (CSV40C, STR-CSV40C, and STR-CH2SV40H2C), including mean diameter, zeta-potential, complex condensation, and decondensation, were evaluated. In addition, cellular and nuclear uptake of plasmid DNA (pDNA) and gene expression in COS7 and dendritic cells (JAWS II) were determined. As a result, C and STR enhanced formation of a smaller and more stable nano-complex with pDNA based on ionic interactions, the disulfide linkage and hydrophobic interactions. STR-CSV40C and STR-CH2SV40H2C had significantly higher cellular uptake ability and transfection efficiency than SV40 and CSV40C. In particular, STR-CH2SV40H2C had higher nuclear uptake and gene expression efficiency than STR-CSV40C. Furthermore, STR-CH2SV40H2C could deliver pDNA to the nuclei and had high gene expression efficiency in dendritic cells. Our results indicate that STR-CH2SV40H2C is a promising gene delivery system in non- or slow-dividing cells.
authors.affiliation: Department of Pharmaceutical Science, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences.
authors.forename: Takanori
authors.initials: T
authors.lastname: Kanazawa
authors.forename: Mamiko
authors.initials: M
authors.lastname: Yamazaki
authors.forename: Tsunehiko
authors.initials: T
authors.lastname: Fukuda
authors.forename: Yuuki
authors.initials: Y
authors.lastname: Takashima
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Okada
mesh_terms: D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D004247:DNA; D015870:Gene Expression; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics; D051379:Mice; D019913:Nuclear Localization Signals / Q000235:genetics*; D009842:Oligopeptides / Q000235:genetics*; D010957:Plasmids
chemical_list: D019913:Nuclear Localization Signals; D009842:Oligopeptides; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D004247:DNA