title: The novel functional nucleic acid iRed effectively regulates target genes following cytoplasmic delivery by faint electric treatment.
abstract: An intelligent shRNA expression device (iRed) contains the minimum essential components needed for shRNA production in cells, and could be a novel tool to regulate target genes. However, general delivery carriers consisting of cationic polymers/lipids could impede function of a newly generated shRNA via electrostatic interaction in the cytoplasm. Recently, we found that faint electric treatment (fET) of cells enhanced delivery of siRNA and functional nucleic acids into the cytoplasm in the absence of delivery carriers. Here, we examined fET of cells stably expressing luciferase in the presence of iRed encoding anti-luciferase shRNA. Transfection of lipofectamine 2000 (LFN)/iRed lipoplexes showed an RNAi effect, but fET-mediated iRed transfection did not, likely because of the endosomal localization of iRed after delivery. However, fET in the presence of lysosomotropic agent chloroquine significantly improved the RNAi effect of iRed/fET to levels that were higher than those for the LFN/iRed lipoplexes. Furthermore, the amount of lipid droplets in adipocytes significantly decreased following fET with iRed against resistin in the presence of chloroquine. Thus, iRed could be a useful tool to regulate target genes following fET-mediated cytoplasmic delivery with endosomal escape devices.
authors.affiliation: Department of Biophysical Chemistry, Kyoto Pharmaceutical University , Kyoto , Japan.
authors.forename: Mahadi
authors.initials: M
authors.lastname: Hasan
authors.affiliation: Tokushima University Graduate School of Biomedical Sciences , Tokushima , Japan.
authors.forename: Noriko
authors.initials: N
authors.lastname: Tarashima
authors.affiliation: Tokushima University Graduate School of Biomedical Sciences , Tokushima , Japan.
authors.forename: Koki
authors.initials: K
authors.lastname: Fujikawa
authors.affiliation: Department of Biophysical Chemistry, Kyoto Pharmaceutical University , Kyoto , Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Ohgita
authors.affiliation: Department of Biophysical Chemistry, Kyoto Pharmaceutical University , Kyoto , Japan.
authors.forename: Susumu
authors.initials: S
authors.lastname: Hama
authors.affiliation: Tokushima University Graduate School of Biomedical Sciences , Tokushima , Japan.
authors.forename: Tamotsu
authors.initials: T
authors.lastname: Tanaka
authors.affiliation: Department of Biophysical Chemistry, Kyoto Pharmaceutical University , Kyoto , Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Saito
authors.affiliation: Tokushima University Graduate School of Biomedical Sciences , Tokushima , Japan.
authors.forename: Noriaki
authors.initials: N
authors.lastname: Minakawa
authors.affiliation: Tokushima University Graduate School of Biomedical Sciences , Tokushima , Japan.
authors.forename: Kentaro
authors.initials: K
authors.lastname: Kogure
keywords: 211 Scaffold / Tissue engineering / Drug delivery; 30 Bio-inspired and biomedical materials; 4′-thioDNA; 600 Others: Nucleic acids; RNAi effect; cytoplasmic delivery; faint electric treatment; iRed