电穿孔
微量注射
生物
转染
神经系统
细胞生物学
胚胎干细胞
中枢神经系统
胚胎
胚胎发生
解剖
分子生物学
神经科学
基因
遗传学
出处
期刊:Nature Protocols
[Nature Portfolio]
日期:2006-08-01
卷期号:1 (3): 1552-1558
被引量:388
标识
DOI:10.1038/nprot.2006.276
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.
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