Sato Masahiro, Inada Emi, Saitoh Issei, Ohtsuka Masato, Nakamura Shingo, Sakurai Takayuki, Watanabe Satoshi
Section of Gene Expression Regulation, Frontier Science Research Center, Kagoshima University, Kagoshima, Japan.
Biotechnol J. 2013 Nov;8(11):1355-61. doi: 10.1002/biot.201300169. Epub 2013 Sep 6.
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.
胰腺被认为是一个重要的基因治疗靶点，因为该器官是多种高负担疾病的发病部位，包括糖尿病、囊性纤维化和胰腺癌。我们旨在开发一种使用非病毒DNA的高效体内基因递送系统。对成年麻醉的ICR雌性小鼠进行胰腺实质内直接注射含环状质粒pmaxGFP DNA的溶液。注射部位用一对镊子型电极盘夹住，并使用方波发生器进行电穿孔。基因递送一天后观察到注射的胰腺部分内绿色荧光蛋白（GFP）表达。转染一周内GFP表达降至基线水平。超过40 V的电压施加会在电穿孔过程中导致组织损伤。我们证明电穿孔对于胰腺细胞的安全高效转染是有效的。这种向胰腺实质递送基因的新方法可能在胰腺疾病的基因治疗策略以及原位特定基因功能的研究中得到应用。
