Zald P B, Cotter M A, Robertso E S
Department of Microbiology and Immunology, University of Michigan Medical School, Ann Arbor 48109-0934, USA.
Prep Biochem Biotechnol. 2001 Feb;31(1):1-11. doi: 10.1081/PB-100103367.
Traditional electroporation devices use direct current electric fields to stimulate the uptake of oligonucleotides, plasmids, short peptides, and proteins into a variety of cell types. A variation of this widely used technique is now available which relies on radio frequency (RF) electrical pulses. This oscillating type of electrical field reportedly elicits greater uptake of plasmid DNA across the plasma membrane. We evaluated a protocol for RF electroporation of the a human embryonic kidney cell line and a Burkitt's lymphoma (BL) cell line for effeciency of transfection by RF electroporation. The plasmid EGFP, which codes for the widely used fusion protein, enhanced green fluorescent protein (EGFP), was used as a reporter of plasmid uptake after transfections. Transfection efficiency consistently increased approximately 30% from that typically obtained with conventional DC type electroporation and was accompanied by greater survivability of cells. Additionally, in some instances, percent transfection efficiency increased to over 70%. Thus, RF electroporation represents an improved methodology for transfection of human cell lines. Moreover, the RF protocol is simple to incorporate in laboratories already utilizing conventional electroporation devices and techniques.
传统的电穿孔设备使用直流电场来刺激寡核苷酸、质粒、短肽和蛋白质进入多种细胞类型。现在有一种这种广泛使用技术的变体,它依赖于射频(RF)电脉冲。据报道,这种振荡类型的电场能使质粒DNA穿过质膜的摄取量更大。我们评估了一种用于人胚肾细胞系和伯基特淋巴瘤(BL)细胞系的射频电穿孔方案,以确定射频电穿孔的转染效率。编码广泛使用的融合蛋白增强绿色荧光蛋白(EGFP)的质粒EGFP,被用作转染后质粒摄取的报告物。转染效率比传统直流型电穿孔通常获得的效率持续提高约30%,并且细胞的存活率更高。此外,在某些情况下,转染效率百分比提高到了70%以上。因此,射频电穿孔代表了一种用于人细胞系转染的改进方法。此外,射频方案很容易纳入已经在使用传统电穿孔设备和技术的实验室中。