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Rapid production of modified cattle and sheep from spermatid-like stem cells

Yang, L.; Di, A.; Song, L.; Liu, X.; Wu, D.; Wang, S.; Hao, Z.; Bu, L.; Bai, C.; Su, G.; Wei, Z.; Zhang, L.; Liu, Z.; Gao, S.; Li, G.

2024-09-14 bioengineering
10.1101/2024.09.10.612264 bioRxiv
Show abstract

Genome-modified ruminants, such as cattle and sheep, are important germplasm for agricultural applications. Haploid androgenetic stem cells (haSCs) can support the full-term development of embryos upon intracytoplasmic haSCs injection (iCHI) into intact oocytes. Thus, haSCs are invaluable resources for studying economic traits and significantly impact livestock breeding; however, ruminant haSCs have yet to be obtained. Here, we report the derivation of cattle and sheep haSCs using a recombinant FACE medium. These cells display characteristics of formative-state pluripotency and can differentiate into three germ layers, both in vitro and in vivo. Interestingly, the ectopic expression of protamine in haSCs converts their nuclei into a spermatid-like structure and synergistic enhancement of full-term development of iCHI embryos, named as Pro-iCHI approach. Furthermore, gene-modified cattle and sheep can be efficiently produced through the combinatorial use of the CRISPR and Pro-iCHI approach. Overall, Pro-iCHI provides an easy-to-manipulate agrobiotechnology tool for rapidly producing genetically modified livestock.

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