Lirong Xiao, Danian Chen, Naihong Yan
Yi Chuan. 2014 Nov;36(11):1173-8. doi: 10.3724/SP.J.1005.2014.1173.
In vivo electroporation is the process referred to a transient increase in the permeability of cell membranes upon high electric field pulses and delivery of engineered constructs into cells. Compared with the viral-mediated gene transfer system, the in vivo electroporation technique has several advantages, such as safe, high efficiency, rapid, stable and wide application. Thus, this technique has been widely used in the studies of many tissues or organs, including the eye. Here, we report the application of in vivo electroporation in the newborn mouse retina. DNA plasmid of GFP expression vector with high concentration was directly injected into the subretinal space of neonatal mouse pups. The DNA was then transfected into the retinal cells after several pulses of high voltage. Transfected GFP allowed clear visualization of cell morphologies in cryo-sections and the GFP was highly expressed in retinal outer nuclear layer. The results showed that this technique can effectively transfer the genes into retinal cells. In vivo electroporation will be a useful tool for the study of retinal development and function.
体内电穿孔是指在高电场脉冲作用下细胞膜通透性短暂增加，并将工程构建体导入细胞的过程。与病毒介导的基因转移系统相比，体内电穿孔技术具有安全、高效、快速、稳定和应用广泛等优点。因此，该技术已广泛应用于包括眼睛在内的许多组织或器官的研究中。在此，我们报告体内电穿孔在新生小鼠视网膜中的应用。将高浓度的绿色荧光蛋白（GFP）表达载体DNA质粒直接注入新生小鼠幼崽的视网膜下间隙。然后在施加几个高压脉冲后，将DNA转染到视网膜细胞中。转染的GFP使得在冷冻切片中能够清晰地观察细胞形态，并且GFP在视网膜外核层中高表达。结果表明，该技术能够有效地将基因转移到视网膜细胞中。体内电穿孔将成为研究视网膜发育和功能的有用工具。
