期刊
GENE THERAPY
卷 19, 期 3, 页码 312-320出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/gt.2011.97
关键词
electropermeabilization; gene electrotransfer; muscle; tempol; reactive oxygen species
类别
资金
- Slovenian Research Agency [P3-0003]
- French-Slovenian Scientific Cooperation
Electropermeabilization (EP) is an effective method of gene transfer into different tissues. During EP, reactive oxygen species (ROS) are formed, which could affect transfection efficiency. The role of generated ROS and the role of antioxidants in electrotransfer in myoblasts in vitro and in Musculus tibialis cranialis in mice were, therefore, investigated. We demonstrate in the study that during EP of C2C12 myoblasts, ROS are generated on the surface of the cells, which do not induce long-term genomic DNA damage. Plasmid DNA for transfection (pEGFP-N1), which is present outside the cells during EP, neutralizes the generated ROS. The ROS generation is proportional to the amplitude of the electric pulses and can be scavenged by antioxidants, such as vitamin C or tempol. When antioxidants were used during gene electrotransfer, the transfection efficiency of C2C12 myoblasts was statistically significantly increased 1.6-fold with tempol. Also in vivo, the transfection efficiency of M. tibialis cranialis in mice was statistically significantly increased 1.4-fold by tempol. The study indicates that ROS are generated on cells during EP and can be scavenged by antioxidants. Specifically, tempol can be used to improve gene electrotransfer into the muscle and possibly also to other tissues. Gene Therapy (2012) 19, 312-320; doi:10.1038/gt.2011.97; published online 30 June 2011
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