Ronald O. Perelman and Claudia Cohen Center for Reproductive Medicine, Weill Cornell Medicine, New York, NY 10065, USA.
Department of Medicine, Weill Cornell Medical College, New York, NY 10065, USA.
CRISPR J. 2021 Feb;4(1):147-154. doi: 10.1089/crispr.2020.0074. Epub 2021 Feb 9.
Mice derived entirely from embryonic stem (ES) cells can be generated through tetraploid complementation. Although XY male ES cell lines are commonly used in this system, occasionally, monosomic XO female mice are produced through spontaneous Y chromosome loss. Here, we describe an efficient method to obtain monosomic XO ES cells by CRISPR-Cas9-mediated deletion of the Y chromosome, allowing generation of female clonal mice by tetraploid complementation. The monosomic XO female mice are viable and able to produce normal male and female offspring. Direct generation of clonal mice in both sexes can significantly accelerate the production of complex genetically modified mouse models.
完全来源于胚胎干细胞（ES）的小鼠可以通过四倍体互补来生成。虽然在该系统中通常使用 XY 雄性 ES 细胞系，但偶尔也会通过自发的 Y 染色体丢失产生单体性 XO 雌性小鼠。在这里，我们描述了一种通过 CRISPR-Cas9 介导的 Y 染色体缺失来获得单体性 XO ES 细胞的有效方法，允许通过四倍体互补生成雌性克隆小鼠。单体性 XO 雌性小鼠具有活力，能够产生正常的雄性和雌性后代。直接生成两性克隆小鼠可以显著加速复杂基因修饰小鼠模型的生产。
