《生命科学》 2010, 22(2): 202-206
摘 要:
摘 要:随着科学研究的不断深入及临床治疗的需要,人们对转基因动物的需求越来越大;但是传统的转基因动物技术大多操作复杂、成本高、效率低,从而限制了转基因技术的广泛应用。利用雄性生殖细胞作为载体介导外源基因导入受精卵来建立转基因动物具有操作简便、经济、易于推广的优点,发展前景广阔。该文就利用雄性生殖细胞建立转基因动物的发展历程和方法进行系统的阐述和分析。从利用精子和精原干细胞携带外源DNA两个方向展开,分别分析和评价了恒温共孵育法、脂质体介导法、电穿孔法、胞浆内单精子注射法、输精管注射法、体外转染精原干细胞法以及体内转染精原干细胞法七种实验设计方法。
关键词:雄性生殖细胞;转基因动物;雄性生殖细胞载体法
Abstract:
Abstract: Along with the development of research and the demand of clinical therapy, the number of transgenic animal needed by people is much more than before. However, most of the traditional techniques for producing transgenic animal were quite complicate, high cost and little effective, which limits their extensive application. Producing transgenic animal by using male germ cell as a vector to introduce the exogenous DNA into zygote has the virtue of simple to handle, affordable and easy to promote, which has a much brighter development prospect. In this review, we systematically expatiated on the course and method of producing transgenic animal by male germ cell and expounded from two directions: one is using the spermatozoa as a vector to introduce the exogenous DNA into zygote; the other is using the spermatogonial stem cell as vector. The introduction and assessment about seven protocols were given, i.e. incubation of sperm and exogenous DNA, lipisome-mediated sperm and exogenous DNA combination, electroporation; intracytoplasmic sperm injection, transfecting mature sperm in vivo by microinjection into vas deferens, transfecting spermatogonial stem cells in vivo and transfecting spermatogonial stem cells in vitro.
Key words: male germ cell; transgenic animal; male germ cell-mediated gene transduction