Real time electroporation control for accurate and safe in vivo non-viral gene therapy
Real time electroporation control for accurate and safe in vivo non-viral gene therapy
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DOI：
10.1016/j.bioelechem.2006.11.001
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发表时间：
2007-05-01
影响因子：
5
通讯作者：
Mir, Lluis M.
中科院分区：
文献类型：
--
作者：
Cukjati, David;Batiuskaite, Danute;Mir, Lluis M.
体内细胞电穿孔是DNA电转移的基础，DNA电转移是一种使用裸DNA进行非病毒基因治疗的有效方法。电脉冲有两个作用，使靶细胞质膜透化，并通过电泳将 DNA 输送至或穿过透化膜。为了有效的电转移，必须进行可逆的、无损的靶细胞透化。我们报告了在脉冲传递过程中监测体内细胞电穿孔的可能性，并在脉冲开始后几微秒内实时调整电场强度，以确保程序的有效性和安全性。详细阐述了控制算法，在原型设备中实施，并在将荧光素酶基因电转移到小鼠肌肉中进行了测试。在基因转移实验中，受控脉冲可以保护组织并产生高水平的荧光素酶，其中未校正的过度施加电压会导致强烈的肌肉损伤和荧光素酶基因表达的连续丧失。 (c) 2006 Elsevier B.V. 保留所有权利。
In vivo cell electroporation is the basis of DNA electrotransfer, an efficient method for non-viral gene therapy using naked DNA. The electric pulses have two roles, to permeabilize the target cell plasma membrane and to transport the DNA towards or across the permeabilized membrane by electrophoresis. For efficient electrotransfer, reversible undamaging target cell permeabilization is mandatory. We report the possibility to monitor in vivo cell electroporation during pulse delivery, and to adjust the electric field strength on real time, within a few microseconds after the beginning of the pulse, to ensure efficacy and safety of the procedure. A control algorithm was elaborated, implemented in a prototype device and tested in luciferase gene electrotransfer to mice muscles. Controlled pulses resulted in protection of the tissue and high levels of luciferase in gene transfer experiments where uncorrected excessive applied voltages lead to intense muscle damage and consecutive loss of luciferase gene expression. (c) 2006 Elsevier B.V. All rights reserved.