Generating single-sex litters: development of CRISPR-Cas9 genetic tools to produce all-male offspring

C. Douglas, V. Maciulyte, J. Zohren, D. M. Snell, O. A. Ojarikre, P. J. Ellis and J. M. A. Turner,  bioRxiv,  2020.09.07.285536. 2020.

Animals are extremely useful genetic tools in science and global resources in agriculture. However, a single sex is often required in surplus, and current genetic methods for producing all-female or all-male litters are inefficient. Using the mouse as a model, we developed a synthetic, two-part bicomponent strategy for generating all-male litters. We achieved this using CRISPR-Cas9 genome editing technology to generate large stable knock-ins on the autosomes and X chromosome. The bicomponent system functions via the sex-specific co-inheritance of a Cas9 transgene and an sgRNA transgene targeting the essential Topoisomerase 1 gene. This technology proved to be highly efficient in generating on-target mutations, resulting in embryonic lethality of the target sex. Our study is the first to successfully generate all-male mammalian litters using a CRISPR-Cas9 bicomponent system and provides great strides towards generating single-sex litters for laboratory or agricultural research.Competing Interest StatementThe authors have declared no competing interest.

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