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Viable semi-cloned mice derived from androgenetic haploid stem cells lacking DNA methylation [RNA-seq]

ABSTRACT: Viable semi-cloned mice derived from androgenetic haploid stem cells lacking DNA methylation [RNA-seq]

PROVIDER: PRJNA1239178 | ENA |

REPOSITORIES: ENA

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Viable semi-cloned mice derived from androgenetic haploid stem cells lacking DNA methylation [TAPS]

Project description:Among all mammalian cell types, sperm exhibit the highest level of DNA methylation, with approximately 70–80% of CpG sites being methylated. The role of this paternal hypermethylation in embryonic developmental competence, aside from the well-characterized H19-DMR and Dlk1-Dio3 intergenic germline DMR (IG-DMR) loci, remains largely unexplored. In mouse male germ cells, the lack of DNA methyltransferase (Dnmt) causes a loss of methylation, leading to meiotic catastrophe and infertility. To circumvent this limitation, we used sperm-like androgenetic haploid embryonic stem cells (AG-haESCs) for oocyte injection to produce offspring. Using CRISPR/Cas9, we effectively inactivated Dnmt1, Dnmt3a and Dnmt3b in AG-haESCs and later reactivated them in the resulting embryos, overcoming Dnmt haploinsufficiency and enabling the generation of viable offspring from methylation-deficient haploid cells. In the offspring embryos, the paternal genome derived from Dnmt-inactivated AG-haESCs rapidly reacquired methylation and restored a methylome in post-implantation embryos comparable to that of wild-type AG-haESC counterparts. These semi-cloned embryos could develop to term and survived to adulthood, exhibiting near-normal morphological and physiological parameters. These findings indicate that the functional significance of paternal genome methylation is mainly restricted to the imprinted loci Igf2-H19 and Dlk1-Dio3, while methylation of the paternal genome elsewhere appears dispensable for normal development.

2025-07-31 | GSE292346 | GEO

Viable semi-cloned mice derived from androgenetic haploid stem cells lacking DNA methylation [RNA-seq]

2025-07-31 | GSE292523 | GEO

Viable semi-cloned mice derived from androgenetic haploid stem cells lacking DNA methylation [bisulfite-Seq]

2025-07-31 | GSE293292 | GEO

Viable semi-cloned mice derived from androgenetic haploid stem cells lacking DNA methylation [TAPS]

Project description:Viable semi-cloned mice derived from androgenetic haploid stem cells lacking DNA methylation [TAPS]

| PRJNA1238204 | ENA

Viable semi-cloned mice derived from androgenetic haploid stem cells lacking DNA methylation [bisulfite-Seq]

Project description:Viable semi-cloned mice derived from androgenetic haploid stem cells lacking DNA methylation [bisulfite-Seq]

| PRJNA1243927 | ENA

Intrinsic de novo DNA methylation in early embryos enables the generation of semi-cloned mice from methylation-deficient androgenetic haploid stem cells

Project description:This SuperSeries is composed of the SubSeries listed below. Abstract Among all mammalian cell types, sperm cells exhibit one of the highest levels of DNA methylation, with ~80% of CpG sites being methylated. The role of this sperm hypermethylation in offspring development, aside from the methylation at the well-characterized H19-DMR and Dlk1-Dio3 intergenic germline DMR (IG-DMR) loci, remains largely unexplored. In mouse germ cells, removal of DNA methylation by deleting methyltransferase (Dnmt) genes causes meiotic catastrophe and infertility. To circumvent this limitation, we inactivated them instead in androgenetic haploid embryonic stem cells (AG-haESCs) lacking H19-DMR and IG-DMR to remove methylation prior to oocyte injection, and subsequently reactivated them in the resulting embryos during cleavage, to ensure Dnmt sufficiency. This strategy enabled the generation of viable offspring from unmethylated paternally-derived haploid cells. In the resulting embryos, the paternal genome rapidly reacquired methylation and was comparable to wild-type in post-implantation embryos. These semi-cloned embryos could develop to term and survived to adulthood, exhibiting near-normal morphological and physiological parameters. These findings indicate that the essential hereditary function of the paternal genome methylation is mainly restricted to the imprinted loci Igf2-H19 and Dlk1-Dio3, while methylation elsewhere appears dispensable for subsequent development, as de novo methylation activity intrinsic to the early embryo ensures epigenetic reprogramming for offspring development.

2025-07-31 | GSE292524 | GEO

Reduced diploidization and survival improvement of semi-cloned mice produced from androgenetic haploid embryonic stem cells

Project description:We found that the overall DNA methylation and hydroxymethylation in AG-haESCs are extremely low, and the downregulation of both de novo methyltransferase Dnmt3b and methylation maintenance enzyme Dnmt1was discovered responsible for this DNA hypomethylation. Further, our study discovered that the correction of DNA methylation can greatly reduce the incidence of diploidization and further improve the survival of semi-cloned mice produced from AG-haESCs.

2017-12-31 | GSE99588 | GEO

CGH analysis of androgenetic haploid embryonic stem cells

Project description:Comparative genomic hybridization (CGH) analysis of androgenetic haploid embryonic stem cells was done with 129/Sv male mouse kidney genomic DNA as reference. The results revealed no consistent copy number variations (CNVs) among the examined cell lines (AH129-5, AH129-N1 and AH129-NC1), except for the global difference on the X chromosome to the male control.

2012-08-30 | GSE39390 | GEO

Mouse Androgenetic Haploid Embryonic Stem Cells Support Targeted Gene Transmission to Progeny

Project description:This SuperSeries is composed of the following subset Series: GSE35785: mRNA expression data from AG-haESC, E14 and MEF GSE35786: CGH analysis of AG-haESCs (androgenetic haploid embryonic stem cells) Refer to individual Series

2012-04-26 | E-GEOD-35787 | biostudies-arrayexpress

Intrinsic de novo DNA methylation in early embryos enables the generation of semi-cloned mice from methylation-deficient androgenetic haploid stem cells

Project description:Intrinsic de novo DNA methylation in early embryos enables the generation of semi-cloned mice from methylation-deficient androgenetic haploid stem cells

| PRJNA1239091 | ENA

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