Project description: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.
Project description: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.
Project description: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.
Project description:Haploid androgenetic embryonic stem cells (haESCs) can produce live mice, thus have potential for genetic manipulation. However, practical application is limited by self-diploidization, loss-of-imprint, low-birth-rate, and livestock haESCs have yet to be achieved. Here we show that combination of FACE medium supports establishment of durable haESCs from cattle and sheep that overcome above problems. Multi-omics and functional analysis of FACE-haESCs showed that at a naive-to-primed intermediate pre-gastrulation formative state of pluripotency. Moreover, transient protamine overexpression remodels ruminant haESCs nuclei into spermatid-like structures concomitant with removal of ICHI barrier, resulting blastocyst rate of Pro-ICHI similar to that of in vitro fertilization (IVF) embryos. As a proof-of-concept, we also developed a novel Pro-ICHI::ePE system to fast produce trait-improved cattle with exogene-free effects.
Project description:Haploid androgenetic embryonic stem cells (haESCs) can produce live mice, thus have potential for genetic manipulation. However, practical application is limited by self-diploidization, loss-of-imprint, low-birth-rate, and livestock haESCs have yet to be achieved. Here we show that combination of FACE medium supports establishment of durable haESCs from cattle and sheep that overcome above problems. Multi-omics and functional analysis of FACE-haESCs showed that at a naive-to-primed intermediate pre-gastrulation formative state of pluripotency. Moreover, transient protamine overexpression remodels ruminant haESCs nuclei into spermatid-like structures concomitant with removal of ICHI barrier, resulting blastocyst rate of Pro-ICHI similar to that of in vitro fertilization (IVF) embryos. As a proof-of-concept, we also developed a novel Pro-ICHI::ePE system to fast produce trait-improved cattle with exogene-free effects.