title: Cytoplasmic gene expression system enhances the efficiency of cationic liposome-mediated in vivo gene transfer into mouse brain.
abstract: Development of methodologies for gene transfer into the central nervous system (CNS) is important for fundamental research as well as clinical studies for gene therapy. Cationic liposomes (CL) are attractive vectors because of their safety and ease of use. However, to date only low rates of success have been reported. We succeeded in obtaining high transfection efficiencies into the newborn mouse brain in vivo by CL and a cytoplasmic gene expression system based on T7 RNA polymerase and T7 RNA polymerase- and the luciferase-gene with the T7 promoter sequence. This system showed an efficiency rate 2 orders of magnitude higher than the standard system, which used CL and luciferase genes with a Rous sarcoma virus promoter, pRSVL. In addition, in vitro experiments using LLCMK2 cells showed that cytoplasmic gene expression occurred rapidly (within 6 h) after transfection. In contrast, pRSVL required 24-48 h for induction of luciferase expression. Our results suggest that the cytoplasmic gene expression system is useful for gene delivery into the CNS.
authors.affiliation: Faculty of Pharmaceutical Science, Osaka University, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Mizuguchi
authors.forename: T
authors.initials: T
authors.lastname: Nakagawa
authors.forename: Y
authors.initials: Y
authors.lastname: Morioka
authors.forename: S
authors.initials: S
authors.lastname: Imazu
authors.forename: M
authors.initials: M
authors.lastname: Nakanishi
authors.forename: T
authors.initials: T
authors.lastname: Kondo
authors.forename: T
authors.initials: T
authors.lastname: Hayakawa
authors.forename: T
authors.initials: T
authors.lastname: Mayumi
mesh_terms: D000818:Animals; D001921:Brain / Q000201:enzymology / Q000378:metabolism*; D002460:Cell Line; D003593:Cytoplasm / Q000235:genetics / Q000378:metabolism*; D012321:DNA-Directed RNA Polymerases / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D007668:Kidney; D008081:Liposomes; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D051379:Mice; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D014162:Transfection / Q000235:genetics; D014764:Viral Proteins
chemical_list: D008081:Liposomes; D014764:Viral Proteins; D008156:Luciferases; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases