CONTROL BY PULSE PARAMETERS OF ELECTRIC FIELD-MEDIATED GENE-TRANSFER IN MAMMALIAN-CELLS
CONTROL BY PULSE PARAMETERS OF ELECTRIC FIELD-MEDIATED GENE-TRANSFER IN MAMMALIAN-CELLS
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DOI：
10.1016/s0006-3495(94)80805-7
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发表时间：
1994-02-01
影响因子：
3.4
通讯作者：
TEISSIE, J
中科院分区：
文献类型：
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作者：
WOLF, H;ROLS, MP;TEISSIE, J
哺乳动物细胞中电场介导的基因转移（电转化）取决于脉冲条件（电场强度、脉冲持续时间、脉冲数）。通过对中国仓鼠卵巢细胞中氯苯乙酰转移酶和β -半乳糖苷酶活性的瞬时表达，系统地研究了这些参数的影响。诱导细胞膜可逆渗透的脉冲条件不足以诱导基因转移。如果质粒要穿过细胞膜进入细胞质，那么在电脉冲期间质粒必须存在。只有被外场带到渗透状态的细胞膜的局部部分是合格的。脉冲持续时间在传输的大小中起着关键作用。电场在细胞膜和质粒之间引起复杂的反应，这种反应通过电泳力积聚在细胞界面上。这导致质粒的插入，然后质粒可以穿过膜。
Electric field-mediated gene transfer in mammalian cells (electrotransformation) depends on the pulsing conditions (field intensity, pulse duration, number of pulses). The effect of these parameters was systematically investigated using the transient expression of the chloramphenycol acetyltransferase and the beta-galactosidase activities in Chinese hamster ovary cells. Pulsing conditions inducing reversible permeabilization of the cell plasma membrane are not sufficient to induce gene transfer. The plasmid must be present during the electric pulse if it is to be transferred across the membrane into the cytoplasm. Only the localized part of the cell membrane brought to the permeabilized state by the external field is competent. Pulse duration plays a key role in the magnitude of the transfer. The field induces a complex reaction between the membrane and the plasmid that is accumulated at the cell interface by electrophoretic forces. This leads to an insertion of the plasmid, which can then cross the membrane.