Generation of completely embryonic stem cell-derived mutant mice using tetraploid blastocyst injection
Generation of completely embryonic stem cell-derived mutant mice using tetraploid blastocyst injection
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DOI：
10.1016/s0925-4773(97)00655-2
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发表时间：
1997-03
影响因子：
2.6
通讯作者：
Zhao-Qi Wang;F. Kiefer;P. Urbánek;E. Wagner
中科院分区：
文献类型：
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作者：
Zhao-Qi Wang;F. Kiefer;P. Urbánek;E. Wagner
胚胎干细胞（ES）为在小鼠中产生专门设计的突变提供了独特的工具。在这里，我们描述了一种替代的方法，对产生的小鼠是完全来自ES细胞（ES小鼠），通过葡萄糖磷酸异构酶（GPI）分析判断，没有事先通过种系。通过将野生型和突变型ES细胞注射到四倍体囊胚中，产生了存活的和可育的ES小鼠，这表明ES细胞的全能性不受长期培养和体外实验操作的影响。当使用通过靶向插入或基因捕获方法引入lacZ报告基因的ES细胞克隆时，ES胎儿中lacZ基因的表达模式与嵌合小鼠繁殖产生的胎儿相同。因此，这种技术可以被认为是一种有用的和快速的方法来产生胚胎和小鼠直接从胚胎干细胞携带预定的遗传变化，并提供了许多应用的分子遗传学和发育生物学的研究。
Embryonic stem (ES) cells provide a unique tool for producing specifically designed mutations in mice. Here, we describe an alternative approach toward the generation of mice which are derived completely from ES cells (ES mice), as judged by glucose phosphate isomerase (GPI) analysis, without prior passage through the germline. By injecting wild-type and mutant ES cells into tetraploid blastocysts, viable and fertile ES mice were generated, suggesting that totipotency of ES cells was not affected by long-term culture and experimental manipulation in vitro. When ES cell clones harboring a lacZ reporter gene introduced by either targeted insertion or a gene-trap approach were used, the expression pattern of the lacZ gene in ES fetuses was identical to that of fetuses that were derived from breeding of chimeric mice. Thus, this technique can be considered as a useful and rapid approach to produce fetuses and mice directly from ES cells carrying predetermined genetic changes and offers many applications for studies in molecular genetics and developmental biology.