Highly efficient constant-current electroporation increases in vivo plasmid expression
Highly efficient constant-current electroporation increases in vivo plasmid expression
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DOI：
10.1089/dna.2005.24.810
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发表时间：
2005-12-01
影响因子：
3.1
通讯作者：
Draghia-Akli, R
中科院分区：
文献类型：
--
作者：
Khan, AS;Pope, MA;Draghia-Akli, R
电穿孔已被证明是一种有效的技术，可增强 DNA 疫苗和治疗蛋白编码质粒向骨骼肌的递送。然而，恒压技术没有考虑组织的电阻，导致组织损伤、炎症和质粒表达丧失。在本研究中，我们使用软件驱动的恒流电穿孔器将质粒传递给小鼠以及小型和大型猪。记录并分析脉冲期间组织的电压、电流强度和电阻。在这两个物种中都确定了电穿孔的最佳条件，并发现其高度依赖于个体组织的抵抗力。与 1 至 2 岁的猪相比，6 至 10 周龄的猪具有更高的肌肉阻力，但这两个值都比小鼠肌肉的阻力低 4 至 5 倍。在小鼠中，质粒注射和电穿孔之间的最佳电流强度、脉冲长度和滞后时间被确定为 0.1 安培、20 毫秒和 0 秒。电极之间的电穿孔脉冲模式也影响质粒表达。这些结果表明，需要针对每个不同的物种优化年龄和组织特异性电阻、脉冲模式以及与电穿孔相关的其他变量，以实现最大的质粒表达。
Electroporation has been demonstrated as an effective technique for enhancing the delivery of plasmids coding for DNA vaccines and therapeutic proteins into skeletal muscle. Nevertheless, constant-voltage techniques do not take into account the resistance of the tissue and result in tissue damage, inflammation, and loss of plasmid expression. In the present study, we have used a software-driven constant-current electroporator to deliver plasmids to mice and small and large pigs. The voltage, amperage, and resistance of the tissue during pulses were recorded and analyzed. Optimal conditions of electroporation were identified in both species, and found to be highly dependent on the individual tissue resistance. Six- to 10-week-old pigs had higher muscle resistance compared to 1- to 2-year-old pigs, but both values were four to five times lower than the resistance of the mouse muscle. In mice, optimum amperage, pulse length, and lag time between plasmid injection and electroporation were identified to be 0.1 Amps, 20 msec and 0 sec. The electroporation pulse pattern among the electrodes also affected plasmid expression. These results indicate that age- and tissue-specific resistance, pulse pattern, and other variables associated with the electroporation need to be optimized for each separate species to achieve maximum plasmid expression.