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基于微电机的哺乳动物细胞局部电穿孔和基因转染。

Micromotor-based localized electroporation and gene transfection of mammalian cells.

机构信息

Faculty of Mechanical Engineering, Micro-, and Nanofluidics Laboratory, Technion - Israel Institute of Technology, Haifa 32000, Israel.

Technion Rappaport Integrated Cancer Center, the Rappaport Faculty of Medicine and Research Institute, Technion-Israel Institute of Technology, Haifa 3109601, Israel.

出版信息

Proc Natl Acad Sci U S A. 2021 Sep 21;118(38). doi: 10.1073/pnas.2106353118.

Abstract

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.

摘要

在这里,我们研究了使用磁性和电力驱动的金属电介质混合微电机对哺乳动物细胞进行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,所提出的微电机有望相对于标准电穿孔增加可逆电穿孔的产率,因为只有细胞膜的一小部分(与微电机接触)受到影响。与将样品中所有细胞的整个细胞膜进行电穿孔的方法不同,所提出的微电机可以通过磁导向对其捕获和运输的目标细胞进行局部、空间精确的电穿孔。为了最小化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,使用磁推进来接近靶细胞的紧邻区域,然后激活短时间的电动推进以实现与细胞接触,随后进行电穿孔。除了局部注射荧光染料分子外,我们还证明由于在电穿孔步骤之前,质粒在詹纳斯粒子和附着的细胞之间的介电泳积聚,微电机可以增强悬浮细胞中质粒的引入。在这里,我们选择了一种不同的策略,涉及许多自主推进、无需外部导向的微电机同时操作,并以自主的方式与细胞配对。局部电穿孔的悬浮细胞被认为很难转染,但也显示出表达转染基因,这对分子生物学研究具有重要意义。

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