Site-targeted non-viral gene delivery by direct DNA injection into the pancreatic parenchyma and subsequent in vivo electroporation in mice |
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| Authors: | Prof Masahiro Sato Emi Inada Issei Saitoh Masato Ohtsuka Shingo Nakamura Takayuki Sakurai Satoshi Watanabe |
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| Institution: | 1. Section of Gene Expression Regulation, Frontier Science Research Center, Kagoshima University, Kagoshima, Japan;2. Department of Pediatric Dentistry, Graduate School of Medical and Dental Sciences, Kagoshima University, Kagoshima, Japan;3. Division of Pediatric Dentistry, Graduate School of Medical and Dental Sciences, Niigata University, Niigata, Japan;4. Division of Basic Molecular Science and Molecular Medicine, School of Medicine, Tokai University, Kanagawa, Japan;5. Department of Surgery, National Defense Medical College, Saitama, Japan;6. Department of Organ Regeneration, Graduate School of Medicine, Shinshu University, Nagano, Japan;7. Animal Genome Research Unit, National Institute of Agrobiological Sciences, Ibaraki, Japan |
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| Abstract: | The pancreas is considered an important gene therapy target because the organ is the site of several high burden diseases, including diabetes mellitus, cystic fibrosis, and pancreatic cancer. We aimed to develop an efficient in vivo gene delivery system using non-viral DNA. Direct intra-parenchymal injection of a solution containing circular plasmid pmaxGFP DNA was performed on adult anesthetized ICR female mice. The injection site was sandwiched with a pair of tweezer-type electrode disks, and electroporated using a square-pulse generator. Green fluorescent protein (GFP) expression within the injected pancreatic portion was observed one day after gene delivery. GFP expression reduced to baseline within a week of transfection. Application of voltages over 40 V resulted in tissue damage during electroporation. We demonstrate that electroporation is effective for safe and efficient transfection of pancreatic cells. This novel gene delivery method to the pancreatic parenchyma may find application in gene therapy strategies for pancreatic diseases and in investigation of specific gene function in situ. |
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| Keywords: | Gene delivery Gene therapy In vivo electroporation Pancreas Site-targeted transfection |
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