Mir Lluis M, Moller Pernille H, André Franck, Gehl Julie
Laboratory of Vectorology and Gene Transfer, UMR 8121 CNRS Institut Gustave-Roussy, F-94805 Villejuif Cédex, France.
Adv Genet. 2005;54:83-114. doi: 10.1016/S0065-2660(05)54005-7.
Electroporation designates the use of electric pulses to transiently permeabilize the cell membrane. It has been shown that DNA can be transferred to cells through a combined effect of electric pulses causing (1) permeabilization of the cell membrane and (2) an electrophoretic effect on DNA, leading the polyanionic molecule to move toward or across the destabilized membrane. This process is now referred to as DNA electrotransfer or electro gene transfer (EGT). Several studies have shown that EGT can be highly efficient, with low variability both in vitro and in vivo. Furthermore, the area transfected is restricted by the placement of the electrodes, and is thus highly controllable. This has led to an increasing use of the technology to transfer reporter or therapeutic genes to various tissues, as evidenced from the large amount of data accumulated on this new approach for non-viral gene therapy, termed electrogenetherapy (EGT as well). By transfecting cells with a long lifetime, such as muscle fibers, a very long-term expression of genes can be obtained. A great variety of tissues have been transfected successfully, from muscle as the most extensively used, to both soft (e.g., spleen) and hard tissue (e.g., cartilage). It has been shown that therapeutic levels of systemically circulating proteins can be obtained, opening possibilities for using EGT therapeutically. This chapter describes the various aspects of in vivo gene delivery by means of electric pulses, from important issues in methodology to updated results concerning the electrotransfer of reporter and therapeutic genes to different tissues.
电穿孔是指利用电脉冲使细胞膜瞬时通透化。研究表明，DNA可通过电脉冲的联合作用转移至细胞，这些电脉冲会导致：（1）细胞膜通透化；（2）对DNA产生电泳效应，使多阴离子分子移向或穿过不稳定的细胞膜。此过程现被称为DNA电转染或电基因转移（EGT）。多项研究表明，EGT效率很高，在体外和体内的变异性都很低。此外，转染区域受电极位置限制，因此具有高度可控性。这使得该技术越来越多地用于将报告基因或治疗性基因转移至各种组织，大量关于这种非病毒基因治疗新方法（也称为电基因治疗，即EGT）的数据积累就证明了这一点。通过转染寿命长的细胞，如肌纤维，可实现基因的长期表达。从使用最广泛的肌肉到软组织（如脾脏）和硬组织（如软骨），多种组织已成功实现转染。研究表明，可获得全身循环蛋白的治疗水平，这为EGT的治疗应用开辟了可能性。本章描述了通过电脉冲进行体内基因递送的各个方面，从方法学中的重要问题到报告基因和治疗性基因向不同组织电转染的最新结果。
