International Journal of Molecular Sciences | |
Intravenous Delivery of piggyBac Transposons as a Useful Tool for Liver-Specific Gene-Switching | |
Satoshi Watanabe1  Masato Ohtsuka2  Naoko Ando3  Masayuki Ishihara3  Shingo Nakamura3  Masahiro Sato4  | |
[1] Animal Genome Unit, Institute of Livestock and Grassland Science, National Agriculture and Food Research Organization (NARO), 2 Ikenodai, Tsukuba, Ibaraki 305-0901, Japan;Division of Basic Medical Science and Molecular Medicine, School of Medicine, Tokai University, Kanagawa 259-1193, Japan;Division of Biomedical Engineering, National Defense Medical College Research Institute, Saitama 359-8513, Japan;Section of Gene Expression Regulation, Frontier Science Research Center, Kagoshima University, Kagoshima 890-8544, Japan; | |
关键词: Cre/loxP; diphtheria toxin-A chain; EGFP; hepatic disorder; hydrodynamics-based gene delivery; in vivo gene delivery; liver; piggyBac transposon; | |
DOI : 10.3390/ijms19113452 | |
来源: DOAJ |
【 摘 要 】
Hydrodynamics-based gene delivery (HGD) is an efficient method for transfecting plasmid DNA into hepatocytes in vivo. However, the resulting gene expression is transient, and occurs in a non-tissue specific manner. The piggyBac (PB) transposon system allows chromosomal integration of a transgene in vitro. This study aimed to achieve long-term in vivo expression of a transgene by performing hepatocyte-specific chromosomal integration of the transgene using PB and HGD. Using this approach, we generated a novel mouse model for a hepatic disorder. A distinct signal from the reporter plasmid DNA was discernible in the murine liver approximately two months after the administration of PB transposons carrying a reporter gene. Then, to induce the hepatic disorder, we first administered mice with a PB transposon carrying a CETD unit (loxP-flanked stop cassette, diphtheria toxin-A chain gene, and poly(A) sites), and then with a plasmid expressing the Cre recombinase under the control of a liver-specific promoter. We showed that this system can be used for in situ manipulation and analysis of hepatocyte function in vivo in non-transgenic (Tg) animals.
【 授权许可】
Unknown