Moreadith R W, Radford N B
Molecular Cardiology Laboratories, University of Texas Southwestern Medical Center, Dallas 75235-8573, USA.
J Mol Med (Berl). 1997 Mar;75(3):208-16. doi: 10.1007/s001090050105.
The development of transgenic technology, whereby genes (or mutations) can be stably introduced into the germline of experimental mammals, now allows investigators to create mice of virtually any genotype and to assess the consequences of these mutations in the context of a developing and intact mammal. In contrast to traditional "gain-of-function" mutations, typically created by microinjection of the gene of interest into the one-celled zygote, gene targeting via homologous recombination in pluripotential embryonic stem cells allows one to modify precisely the gene of interest. The purpose of this review is to introduce the reader to the history of development of embryonic stem cell technology, the current methods employed to create "knock-out" mice, and the application of these methods to solve problems in biology. While the technology promises to provide enormous insight into mammalian development genetics, our desire is that this review will stimulate the application of gene targeting in embryonic stem cells to begin to unravel problems in complex regulatory pathways, specifically intermediary metabolism and physiology.
转基因技术的发展，使基因（或突变）能够稳定地导入实验哺乳动物的种系，现在研究人员可以创造出几乎任何基因型的小鼠，并在发育完整的哺乳动物体内评估这些突变的后果。与传统的“功能获得性”突变不同，后者通常是通过将感兴趣的基因显微注射到单细胞受精卵中产生的，而通过多能胚胎干细胞中的同源重组进行基因靶向，能让人们精确修饰感兴趣的基因。这篇综述的目的是向读者介绍胚胎干细胞技术的发展历程、目前用于创建“基因敲除”小鼠的方法，以及这些方法在解决生物学问题中的应用。虽然这项技术有望为哺乳动物发育遗传学提供巨大的见解，但我们希望这篇综述能激发在胚胎干细胞中应用基因靶向技术，从而开始解决复杂调控途径中的问题，特别是中间代谢和生理学问题。
