Site-targeted non-viral gene delivery by direct DNA injection into the pancreatic parenchyma and subsequent in vivo electroporation in mice.
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.
Section of Gene Expression Regulation, Frontier Science Research Center, Kagoshima University, Kagoshima, Japan. masasato@ms.kagoshima-u.ac.jp.
Masahiro
M
Sato
Emi
E
Inada
Issei
I
Saitoh
Masato
M
Ohtsuka
Shingo
S
Nakamura
Takayuki
T
Sakurai
Satoshi
S
Watanabe
D000818:Animals; D018274:Electroporation / Q000379:methods*; D005260:Female; D049452:Green Fluorescent Proteins / Q000235:genetics; D007267:Injections; D051379:Mice; D008813:Mice, Inbred ICR; D010179:Pancreas / Q000166:cytology / Q000378:metabolism*; D010957:Plasmids / Q000008:administration & dosage* / Q000235:genetics; D014162:Transfection / Q000379:methods*
D049452:Green Fluorescent Proteins
Gene delivery; Gene therapy; In vivo electroporation; Pancreas; Site-targeted transfection