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Rapid generation of gene-edited cattle and sheep from spermatid-like formative haESCs [WGS]

ABSTRACT: 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.

ORGANISM(S): Bos taurus Ovis aries

PROVIDER: GSE250496 | GEO | 2025/08/04

REPOSITORIES: GEO

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Rapid generation of gene-edited cattle and sheep from spermatid-like formative haESCs [WGBS]

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.

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