In vivo electroporation in the embryonic mouse central nervous system

被引:296
|
作者
Saito, Tetsuichiro [1 ]
机构
[1] Chiba Univ, Grad Sch Med, Dept Dev Biol, Chiba 2608670, Japan
关键词
D O I
10.1038/nprot.2006.276
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
This protocol describes a basic method for in vivo electroporation in the nervous system of embryonic mice. Delivery of electric pulses following microinjection of DNA into the brain ventricle or the spinal cord central canal enables efficient transfection of genes into the nervous system. Transfection is facilitated by forceps-type electrodes, which hold the uterus and/or the yolk sac containing the embryo. More than ten embryos in a single pregnant mouse can be operated on within 30 min. More than 90% of operated embryos survive and more than 90% of these survivors express the transfected genes appropriately. Gene expression in neurons persists for a long time, even at postnatal stages, after electroporation. Thus, this method could be used to analyze roles of genes not only in embryonic development but also in higher order function of the nervous system, such as learning.
引用
收藏
页码:1552 / 1558
页数:7
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