| Summary: | 碩士 === 國立臺灣大學 === 獸醫學研究所 === 88 === Abstract
Electroporation(EP) is the transient permeabilization of the plasma membrane by means of short and intense electric pulses. Under optimized condition, electroporation is compatible with cell survival. It provides a direct access into the cytosol to exogenous drugs and genes. Limitations from other gene targeting methods such as low transfection efficiencies, inconsistent reproducibility, safety concerns and cellular toxicity do not exist for the application of electroporation.
The efficiency of in vivo-electro-transfection on gene expression was first evaluated by green fluorescent protein (GFP) gene with fluorescence microscope. The transfer efficiency can be high up to 84.4%. We applied this gene-targeting technology on the study of immunotherapy on mouse model by transfecting mIL-2 gene into established 1ME A.7R.1tumor by in vivo electroporation. Three rounds of electroporations are needed for an effective immuno-stimulation by locally applied mIL-2 genes. The anti-tumor activity can be increased by double gene transfer with both mIL-2 gene and mGM-CSF gene delivered simultaneously.
In conclusion, we have estabilished an novel immunotherapy approach by in vivo gene electroporation. The efficiency and efficacy of targeted gene are discussed. In order to apply this technology on veterinary field, we need to develop a better delivery system for permeabilizing electric pulses to the entire tumor to be treated. Effective local immunity may provide an effective systemic anti-tumor effects.
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