Generation of clonal male and female mice through CRISPR/Cas9-mediated Y chromosome deletion in embryonic stem cells

Mice derived entirely from embryonic stem (ES) cells can be generated in one step through tetraploid complementation. Although XY male ES cell lines are commonly used in this system, occasionally, monosomic XO female all-ES 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 clonal male and female mice by tetraploid complementation. The monosomic XO female mice are viable and are able to produce normal male and female offspring. Direct generation of clonal male and female mice from the same mutant ES cells significantly accelerates the production of complex genetically modified mouse models by circumventing multiple rounds of outbreeding.Competing Interest StatementThe authors have declared no competing interest.View Full Text