Open AccessGeneration of Sex-Reversed Female Clonal Mice via CRISPR-Cas9-Mediated Y Chromosome Deletion in Male Embryonic Stem Cells
Author(s) -
Yiren Qin,
Bokey Wong,
Liangwen Zhong,
Fuqiang Geng,
Luis F. Parada,
Desheng Wen
Publication year - 2021
Publication title -
the crispr journal
Language(s) - English
Resource type - Journals
eISSN - 2573-1602
pISSN - 2573-1599
DOI - 10.1089/crispr.2020.0074
Subject(s) - biology , offspring , crispr , embryonic stem cell , complementation , genetics , cas9 , y chromosome , chromosome , microbiology and biotechnology , gene , phenotype , pregnancy
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.