title: PhiC31 integrase-mediated genomic integration and stable gene expression in the mouse mammary gland after gene electrotransfer.
abstract: BACKGROUND
  PhiC31 integrase is capable of conferring long-term transgene expression in various transfected tissues in vivo. In the present study, we investigated the activity of phiC31 integrase in mouse mammary glands.
  
  
  METHODS
  The normal mouse mammary epithelial cell line HC11 was transfected with FuGENE® HD Transfection Reagent (Roche Diagnostics, Shanghai, China). Transfection of the mouse mammary gland in vivo was performed by electrotransfer. Transgene expression was detected by western blotting and an enzyme-linked immunosorbent assay. Genomic integration and integration at mpsL1 was confirmed by a nested polymerase chain reaction.
  
  
  RESULTS
  An optimal electrotransfer protocol for the lactating mouse mammary gland was attained through investigation of different voltages and pulse durations. PhiC31 integrase mediated site-specific transgene integration in HC11 cells and the mouse mammary gland. In addition, the site-specific integration occurred efficiently at the ‘hot spot’ mpsL1. Co-delivery of PhiC31 integrase enhanced and prolonged transgene expression in the mouse mammary gland.
  
  
  CONCLUSIONS
  The results obtained in the present study show that the use of phiC31 integrase is a feasible and efficient method for high and stable transgene expression in the mouse mammary gland.
authors.forename: Yan
authors.initials: Y
authors.lastname: Luo
authors.forename: Jun
authors.initials: J
authors.lastname: Liu
authors.forename: Yongsheng
authors.initials: Y
authors.lastname: Wang
authors.forename: Jianmin
authors.initials: J
authors.lastname: Su
authors.forename: Yongyan
authors.initials: Y
authors.lastname: Wu
authors.forename: Guangdong
authors.initials: G
authors.lastname: Hu
authors.forename: Mingqing
authors.initials: M
authors.lastname: Gao
authors.forename: Fusheng
authors.initials: F
authors.lastname: Quan
authors.forename: Yong
authors.initials: Y
authors.lastname: Zhang
mesh_terms: D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D018274:Electroporation / Q000379:methods; D018628:Gene Dosage; D015870:Gene Expression; D018014:Gene Transfer Techniques; D056426:Genetic Loci; D005822:Genetic Vectors / Q000235:genetics; D016678:Genome; D059765:Homologous Recombination; D006801:Humans; D019426:Integrases / Q000235:genetics / Q000378:metabolism*; D042361:Mammary Glands, Human / Q000378:metabolism*; D051379:Mice; D014162:Transfection; D019076:Transgenes
chemical_list: D019426:Integrases