Department of Cell and Developmental Biology, University of Colorado Denver, Anschutz Medical Campus, Aurora, CO 80045, USA.
Mamm Genome. 2009 Nov-Dec;20(11-12):734-40. doi: 10.1007/s00335-009-9228-z. Epub 2009 Oct 1.
Genetic manipulation of embryonic stem (ES) cells has been used to produce genetically engineered mice modeling human disorders. Here we describe a novel, additional application: selection for a phenotype of interest and subsequent transmission of that phenotype to a living mouse. We show, for the first time, that a cellular phenotype induced by ENU mutagenesis in ES cells can be transmitted and recapitulated in adult mice derived from these cells. We selected for paraquat-resistant (PQ(R)) ES clones. Subsequent injection of these cells into blastocysts resulted in the production of germline chimeras, from which tail skin fibroblasts exhibited enhanced PQ(R). This trait was also recovered in progeny of the chimera. We avoided PQ toxicity, which blocks the ability to involve the germline, by developing a sib-selection method, one that could be widely applied wherever the selection itself might diminish the pluripotency of the ES cells. Thus, phenotype-driven screens in ES cells are both feasible and efficient in producing intact mouse models for in vivo studies.
胚胎干细胞（ES）的基因操作已被用于产生模拟人类疾病的基因工程小鼠。在这里，我们描述了一种新颖的应用：选择感兴趣的表型，然后将该表型传递给活体小鼠。我们首次表明，由 ES 细胞中 ENU 诱变诱导的细胞表型可以在源自这些细胞的成年小鼠中传递和再现。我们选择百草枯抗性（PQ（R））ES 克隆。随后将这些细胞注射到囊胚中会产生种系嵌合体，从中提取的尾巴皮肤成纤维细胞表现出增强的 PQ（R）。该特征也在嵌合体的后代中恢复。我们通过开发一种同胞选择方法来避免 PQ 毒性，该毒性会阻止种系参与，这种方法可以广泛应用于选择本身可能降低 ES 细胞多能性的任何地方。因此，在 ES 细胞中进行表型驱动的筛选在产生用于体内研究的完整小鼠模型方面既可行又有效。
