Department of Molecular Medicine, University of South Florida, Tampa, FL, USA.
Gene Ther. 2011 May;18(5):496-500. doi: 10.1038/gt.2010.171. Epub 2010 Dec 23.
Electroporation (EP) is a simple in vivo method to deliver normally impermeable molecules, such as plasmid DNA, to a variety of tissues. Delivery of plasmid DNA by EP to a large surface area is not practical because the distance between the electrode pairs, and therefore the applied voltage, must be increased to effectively permeabilize the cell membrane. The design of the multielectrode array (MEA) incorporates multiple electrode pairs at a fixed distance to allow for delivery of plasmid DNA to the skin, potentially reducing the sensation associated with in vivo EP. In this report, we evaluate the effects of field strength and pulse width on transgene expression and duration using a plasmid encoding the luciferase reporter gene delivered by intradermal injection in a guinea pig model followed by EP with the MEA. As expected, the level of luciferase expression increased with the magnitude and duration of the voltage applied. In addition to adjusting transgene expression levels by altering fielding strength, levels could also be controlled by adjusting the plasmid dose. Our results indicate that the design of the MEA is a viable option for cutaneous plasmid DNA delivery by in vivo EP to a large surface area.
电穿孔(EP)是一种将通常不可渗透的分子(如质粒 DNA)递送到各种组织的简单体内方法。由于电极对之间的距离(因此应用的电压)必须增加才能有效地使细胞膜穿孔,因此通过 EP 将质粒 DNA递送到大面积是不切实际的。多电极阵列(MEA)的设计采用固定距离的多个电极对,允许将质粒 DNA递送到皮肤,从而可能降低与体内 EP 相关的感觉。在本报告中,我们使用通过皮内注射递送至豚鼠模型的编码荧光素酶报告基因的质粒,并随后通过 MEA 进行 EP,评估场强和脉冲宽度对转基因表达和持续时间的影响。正如预期的那样,荧光素酶表达水平随施加电压的幅度和持续时间的增加而增加。除了通过改变场强来调整转基因表达水平外,还可以通过调整质粒剂量来控制水平。我们的结果表明,MEA 的设计是通过体内 EP 将大量皮肤的质粒 DNA 递送到大面积的可行选择。