PhiC31 integrase-mediated genomic integration and stable gene expression in the mouse mammary gland after gene electrotransfer

作者：Luo, Yan; Liu, Jun; Wang, Yongsheng; Su, Jianmin; Wu, Yongyan; Hu, Guangdong; Gao, Mingqing; Quan, Fusheng; Zhang, Yong^*

来源：JOURNAL OF GENE MEDICINE, 2013, 15(10): 356-365.

DOI：10.1002/jgm.2723

• electrotransfer
• gene delivery
• gene expression
• mouse mammary gland
• phiC31 integrase

BackgroundPhiC31 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.
MethodsThe normal mouse mammary epithelial cell line HC11 was transfected with FuGENE (R) 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.
ResultsAn 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.
ConclusionsThe 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.

• 出版日期2013-10
• 单位西北农林科技大学