Rapid production of modified cattle and sheep from spermatid-like stem cells [CUT&Tag]

Haploid androgenetic stem cells (haSCs) are invaluable resources for studying animal traits and greatly impact livestock breeding, but livestock haSCs have yet to be obtained. Here, we report the derivation of cattle and sheep haSCs using the recombined FACE medium, and these cells harbor formative pluripotency features. Notably, knockout cattle and sheep can be obtained upon injection of the protaminized haSCs with targeted gene mutations into intact oocytes. Combination of F, A, C, and E small-molecule supports establishment of bovine haSCs (input, H3K4me3, H3K27me3, PRM-0h, PRM-24h, PRM-48h, PRM-72h) at formative state of pluripotency.