Department of Power Mechanical Engineering, National Tsing Hua University, Hsinchu, Taiwan.
Lab Chip. 2014 Feb 7;14(3):592-601. doi: 10.1039/c3lc51102b. Epub 2013 Dec 9.
Gene transfection is an important technology for various biological applications. The exogenous DNA is commonly delivered into cells by using a strong electrical field to form transient pores in cellular membranes. However, the high voltage required in this electroporation process may cause cell damage. In this study, a dielectrophoretically-assisted electroporation was developed by using light-activated virtual microelectrodes in a new microfluidic platform. The DNA electrotransfection used a low applied voltage and an alternating current to enable electroporation and transfection. Single or triple fluorescence-carrying plasmids were effectively transfected into various types of mammalian cells, and the fluorescent proteins were successfully expressed in live transfected cells. Moreover, the multi-triangle optical pattern that was projected onto a photoconductive layer to generate localized non-uniform virtual electric fields was found to have high transfection efficiency. The developed dielectrophoretically-assisted electroporation platform may provide a simpler system for gene transfection and could be widely applied in many biotechnological fields.
基因转染是各种生物应用的重要技术。通常通过使用强电场在细胞膜中形成瞬时孔来将外源 DNA 递送到细胞中。然而，电穿孔过程中所需的高电压可能会导致细胞损伤。在这项研究中，通过在新的微流控平台中使用光激活虚拟微电极，开发了介电泳辅助电穿孔。DNA 电转染使用低应用电压和交流电来进行电穿孔和转染。单或三荧光载体质粒有效地转染到各种类型的哺乳动物细胞中，并且荧光蛋白在活转染细胞中成功表达。此外，发现投射到光电导层上以产生局部非均匀虚拟电场的多三角形光学图案具有高转染效率。开发的介电泳辅助电穿孔平台可为基因转染提供更简单的系统，并可广泛应用于许多生物技术领域。
