Robertson E, Bradley A, Kuehn M, Evans M
Nature. 1986;323(6087):445-8. doi: 10.1038/323445a0.
Embryonic stem cells isolated directly from mouse embryos can be cultured for long periods in vitro and subsequently repopulate the germ line in chimaeric mice. During the culture period these embryonic cells are accessible for experimental genetic manipulation. Here we report the use of retroviral vectors to introduce exogenous DNA sequences into a stem-cell line and show that these modified cells contribute extensively to the somatic and germ-cell lineages in chimaeric mice. Compared with current methods for manipulation of the mouse genome, this approach has the advantage that powerful somatic-cell genetic techniques can be used to modify and to select cells with germ-line potential, allowing the derivation of transgenic strains with pre-determined genetic changes. We have by this means inserted many proviral vector sequences that provide new chromosomal molecular markers for linkage studies in the mouse and that also may cause insertional mutations.
直接从小鼠胚胎中分离出的胚胎干细胞可在体外长期培养，随后在嵌合小鼠中重新填充生殖系。在培养期间，这些胚胎细胞可用于实验性基因操作。在此，我们报告了使用逆转录病毒载体将外源DNA序列导入干细胞系，并表明这些经过修饰的细胞在嵌合小鼠的体细胞和生殖细胞谱系中广泛发挥作用。与目前操纵小鼠基因组的方法相比，这种方法的优点是可以使用强大的体细胞遗传技术来修饰和选择具有生殖系潜能的细胞，从而获得具有预先确定基因变化的转基因品系。通过这种方式，我们插入了许多前病毒载体序列，这些序列为小鼠的连锁研究提供了新的染色体分子标记，并且也可能导致插入突变。
