Abstract
Known examples of male to female sex reversal in mice are caused by either strain incompatibilities or mutations in genes required for male sex determination. The resultant XY females are often sterile or exhibit very poor fertility. We describe here embryonic stem (ES) cell growth conditions that promote the production of healthy, anatomically normal fertile and fecund female F0 generation mice completely derived from gene-targeted XY male ES cells. The sex reversal is a transient trait that is not transmitted to the F1 progeny. Growth media with low osmolality and reduced sodium bicarbonate, maintained throughout the gene targeting process, enhance the yield of XY females. As a practical application of the induced sex reversal, we demonstrate the generation of homozygous mutant mice ready for phenotypic studies by the breeding of F0 XY females with their isogenic XY male clonal siblings, thereby eliminating one generation of breeding and the associated costs.
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Acknowledgments
We dedicate this work to Roberta Rivi, whose seminal contributions were crucial to the discovery of the XY female phenomenon. We acknowledge the contributions of the entire VelociGene team, in particular, Kate Kudynska and Jade Zhang for phenotype studies, Michael Brown and Joseph Hickey for ES cell microinjections, Alejo Mujica for figure generation and Jeffrey Lee for critical improvements to the manuscript. We thank Jennifer Schmahl for helpful discussions on sex determination.
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11248_2014_9815_MOESM1_ESM.tiff
Sperm defects and reduced testis size of F1 generation XXY and XYY male progeny of XY females. (a) Sperm count. (b) Sperm motility. (c) Testis Length. (d) Testis weight. Each data point represents the measurement for a single mouse. Horizontal bars represent the mean bracketed by SEM. (e) Dissected testes show the smaller size in an XYY male compared with those from an XY male (TIFF 643 kb)
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Kuno, J., Poueymirou, W.T., Gong, G. et al. Generation of fertile and fecund F0 XY female mice from XY ES cells. Transgenic Res 24, 19–29 (2015). https://doi.org/10.1007/s11248-014-9815-y
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DOI: https://doi.org/10.1007/s11248-014-9815-y