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PhiC31 integrase‐mediated genomic integration and stable gene expression in the mouse mammary gland after gene electrotransfer
Author(s) -
Luo Yan,
Liu Jun,
Wang Yongsheng,
Su Jianmin,
Wu Yongyan,
Hu Guangdong,
Gao Mingqing,
Quan Fusheng,
Zhang Yong
Publication year - 2013
Publication title -
the journal of gene medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.689
H-Index - 91
eISSN - 1521-2254
pISSN - 1099-498X
DOI - 10.1002/jgm.2723
Subject(s) - integrase , transgene , transfection , biology , microbiology and biotechnology , mammary gland , genetically modified mouse , gene , genetics , cancer , breast cancer
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. Copyright © 2013 John Wiley & Sons, Ltd.