Foreign gene expression in an organotypic culture of cortical anlage after in vivo electroporation.
Foreign gene expression in an organotypic culture of cortical anlage after in vivo electroporation.
复制标题
体内电穿孔后皮质原基器官型培养物中的外源基因表达。
DOI：
10.1097/00001756-199908020-00018
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发表时间：
1999
期刊：
影响因子：
1.7
通讯作者：
K. Takiguchihayashi
中科院分区：
文献类型：
--
作者：
N. Miyasaka;Y. Arimatsu;K. Takiguchihayashi
通过电穿孔介导的体内基因转移，在大脑皮层原基的器官型培养物中实现了外源基因的高水平表达。将绿色荧光蛋白（GFP）基因的哺乳动物表达质粒注入大鼠胚胎侧脑室。质粒DNA注射后，立即在一对镊子型电极之间对胚胎的头部进行电穿孔。分离皮质原基，并在体外（DIV）保持器官型长达21天。GFP转基因在1 DIV时在神经前体细胞中强烈表达。GFP表达细胞仍然是可检测的，并证明在21 DIV分化成神经元和神经胶质。该系统有望用于大脑皮层发育和功能的分子分析。
A high level of foreign gene expression in organotypic cultures of the cerebral cortical anlage was achieved by electroporation-mediated gene transfer in vivo. A mammalian expression plasmid for green fluorescent protein (GFP) gene was injected into the lateral ventricle of rat embryos. Immediately after the plasmid DNA injection, the head of the embryo was electroporated between a pair of tweezer-type electrodes. The cortical anlage was isolated and maintained organotypically up to 21 days in vitro (DIV). The GFP-transgene was expressed intensely in neural progenitor cells at 1 DIV. GFP-expressing cells were still detectable and were demonstrated to differentiate into neurons and glia at 21 DIV. This system is expected to be useful for molecular analysis of cerebral cortical development and function.