Efficient gene transfer into the embryonic mouse brain using in vivo electroporation
Efficient gene transfer into the embryonic mouse brain using in vivo electroporation
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DOI：
10.1006/dbio.2001.0439
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发表时间：
2001-12-01
影响因子：
2.7
通讯作者：
Nakatsuji, N
中科院分区：
文献类型：
--
作者：
Saito, T;Nakatsuji, N
小鼠遗传操作提供了一个很好的系统，在许多情况下，基因功能的特点。通过转基因、基因敲除和诱变技术已经产生了许多突变体。然而，一个限制是难以以限制性方式（即，空间上和时间上），因为可以限制基因表达的可用增强子和启动子的数量有限。我们已经开发了一种新的方法，通过使用电穿孔将DNA导入子宫内/子宫外胚胎小鼠脑的不同阶段。超过90%的手术胚胎存活下来，其中超过65%的胚胎在大脑的限制区域表达了引入的基因。即使在出生后，电穿孔后6周，表达仍保持不变。荧光蛋白基因的使用清楚地显示了大脑中的神经元形态。此外，有可能将三种不同的DNA载体转染到相同的细胞中。因此，这种方法将是一个强大的工具，以表征基因功能在各种设置，由于其高效率和本地化的基因表达。(C)北京：科学出版社.
Mouse genetic manipulation has provided an excellent system to characterize gene function in numerous contexts. A number of mutants have been produced by using transgenic, gene knockout, and mutagenesis techniques. Nevertheless, one limitation is that it is difficult to express a gene in vivo in a restricted manner (i.e., spatially and temporally), because the number of available enhancers and promoters which can confine gene expression is limited. We have developed a novel method to introduce DNA into in/exo utero embryonic mouse brains at various stages by using electroporation. More than 90% of operated embryos survived, and more than 65% of these expressed the introduced genes in restricted regions of the brain. Expression was maintained even after birth, 6 weeks after electroporation. The use of fluorescent protein genes clearly visualized neuronal morphologies in the brain. Moreover, it was possible to transfect three different DNA vectors into the same cells. Thus, this method will be a powerful tool to characterize gene function in various settings due to its high efficiency and localized gene expression. (C) 2001 Academic Press.