Haas K, Sin W C, Javaherian A, Li Z, Cline H T
Cold Spring Harbor Laboratory, Cold Spring Harbor, NY 11724, USA.
Neuron. 2001 Mar;29(3):583-91. doi: 10.1016/s0896-6273(01)00235-5.
We report an electroporation technique for targeting gene transfer to individual cells in intact tissue. Electrical stimulation through a micropipette filled with DNA or other macromolecules electroporates a single cell at the tip of the micropipette. Electroporation of a plasmid encoding enhanced green fluorescent protein (GFP) into the brain of intact Xenopus tadpoles or rat hippocampal slices resulted in GFP expression in single neurons and glia. In vivo imaging showed morphologies, dendritic arbor dynamics, and growth rates characteristic of healthy cells. Coelectroporation of two plasmids resulted in expression of both proteins, while electroporation of fluorescent dextrans allowed direct visualization of transfer of molecules into cells. This technique will allow unprecedented spatial and temporal control over gene delivery and protein expression.
我们报告了一种电穿孔技术，用于将基因转移靶向完整组织中的单个细胞。通过充满DNA或其他大分子的微吸管进行电刺激，可使微吸管尖端的单个细胞发生电穿孔。将编码增强型绿色荧光蛋白（GFP）的质粒电穿孔导入完整非洲爪蟾蝌蚪的大脑或大鼠海马切片中，导致单个神经元和神经胶质细胞中出现GFP表达。体内成像显示出健康细胞特有的形态、树突分支动态和生长速率。两种质粒的共电穿孔导致两种蛋白质都表达，而荧光葡聚糖的电穿孔则允许直接观察分子向细胞内的转移。这项技术将实现对基因传递和蛋白质表达前所未有的空间和时间控制。
