Micromotor-based localized electroporation and gene transfection of mammalian cells [Engineering],Proceedings of the National Academy of Sciences of the United States of America

Herein, we studied localized electroporation and gene transfection of mammalian cells using a metallodielectric hybrid micromotor that is magnetically and electrically powered. Much like nanochannel-based, local electroporation of single cells, the presented micromotor was expected to increase reversible electroporation yield, relative to standard electroporation, as only a small portion of the cell’s membrane (in contact with the micromotor) is affected. In contrast to methods in which the entire membrane of all cells within the sample are electroporated, the presented micromotor can perform, via magnetic steering, localized, spatially precise electroporation of the target cells that it traps and transports. In order to minimize nonselective electrical lysis of all cells within the chamber, resulting from extended exposure to an electrical field, magnetic propulsion was used to approach the immediate vicinity of the targeted cell, after which short-duration, electric-driven propulsion was activated to enable contact with the cell, followed by electroporation. In addition to local injection of fluorescent dye molecules, we demonstrated that the micromotor can enhance the introduction of plasmids into the suspension cells because of the dielectrophoretic accumulation of the plasmids in between the Janus particle and the attached cell prior to the electroporation step. Here, we chose a different strategy involving the simultaneous operation of many micromotors that are self-propelling, without external steering, and pair with cells in an autonomic manner. The locally electroporated suspension cells that are considered to be very difficult to transfect were shown to express the transfected gene, which is of significant importance for molecular biology research.

中文翻译：

基于微电机的哺乳动物细胞局部电穿孔和基因转染 [工程]

在此，我们使用磁性和电动的金属介电混合微电机研究了哺乳动物细胞的局部电穿孔和基因转染。与基于纳米通道的单细胞局部电穿孔非常相似，相对于标准电穿孔，所提出的微电机有望增加可逆电穿孔产量，因为只有一小部分细胞膜（与微电机接触）受到影响。与样品中所有细胞的整个膜都被电穿孔的方法相比，所提出的微电机可以通过磁力转向对它捕获和运输的靶细胞进行局部、空间精确的电穿孔。为了最大限度地减少由于长时间暴露于电场而导致的腔室内所有细胞的非选择性电裂解，磁力推进被用来接近目标细胞的附近，之后短时间的电驱动推进被激活以与细胞接触，然后进行电穿孔。除了局部注射荧光染料分子外，我们还证明了微电机可以增强质粒向悬浮细胞的引入，因为在电穿孔步骤之前，质粒在 Janus 颗粒和附着的细胞之间存在介电泳积累。在这里，我们选择了一种不同的策略，包括同时运行许多自驱动的微型电机，无需外部转向，并以自主方式与细胞配对。