GERM-LINE TRANSMISSION OF GENES INTRODUCED INTO CULTURED PLURIPOTENTIAL CELLS BY RETROVIRAL VECTOR
GERM-LINE TRANSMISSION OF GENES INTRODUCED INTO CULTURED PLURIPOTENTIAL CELLS BY RETROVIRAL VECTOR
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DOI：
10.1038/323445a0
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发表时间：
1986-10-02
期刊：
影响因子：
64.8
通讯作者：
EVANS, M
中科院分区：
文献类型：
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作者：
ROBERTSON, E;BRADLEY, A;EVANS, M
直接从小鼠胚胎中分离出来的胚胎干细胞可以在体外长期培养，然后在嵌合小鼠中重新繁殖生殖系2、3。在培养期间，这些胚胎细胞可用于实验性基因操作4-6。在这里，我们报道了使用逆转录病毒载体将外源DNA序列引入干细胞系，并表明这些修饰的细胞对嵌合小鼠的体细胞和生殖细胞谱系做出了广泛贡献。与目前的小鼠基因组操作方法相比，该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种系潜力的细胞，从而产生具有预先确定的遗传变化的转基因菌株。我们通过这种方式插入了许多原病毒载体序列，这些序列为小鼠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。
Embryonic stem cells isolated directly from mouse embryos1can be cultured for long periodsin vitroand subsequently repopulate the germ line in chimaeric mice2,3. During the culture period these embryonic cells are accessible for experimental genetic manipulation4–6. 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.