Engineering mice for female-biased progeny without impacting genetic integrity and litter size

Abstract Generating mammalian gametes with a skewed sex ratio has thus far eluded empirical confirmation. The utilization of such genetically engineered organisms would offer the potential to curtail the necessity for culling animals of undesirable sex, mitigate resource wastage, and alleviate superfluous labor burdens. In this study, we introduce a transgenic male mouse lineage, which consistently yields predominantly female progeny (comprising up to ～94% of the total offspring). This accomplishment was made possible by integrating a controllable genetic cassette onto the Y chromosome. The cassette encodes dCas9 and RNA guides that selectively silence a spermatid maturation gene. After the separation of X and Y gametes during meiosis, gametes containing an X chromosome develop normally, while those harboring the engineered Y chromosome, subjected to dCas9 silencing of the spermatid maturation gene, do not mature properly. Indeed, some spermatozoa from the transgenic mice exhibit a unique morphology, associated with the absence of the maturation gene. Notably, the resultant female offspring do not inherit the genetically engineered Y chromosome and are thus not genetically modified. Importantly, the litter size of the transgenic mice remains unchanged compared to the wild type. These findings represent a groundbreaking demonstration of genetic engineering’s potential to yield sex-biased litters of full size without compromising genetic integrity, marking a pioneering advancement in this field of study. biorxiv;2023.11.21.568055v2/FIGA1F1figa1Graphical Abstract.A proof of concept for the first mammal producing sexed semen may revolutionize the way for breeding animals such as cows for the dairy industry and chickens for the eggs industry. One sentence summaryY chromosome manipulation yields mostly female mice, preserving litter size without genetic alteration in females.