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Epigenetic integrity of paternal imprints enhances the developmental potential of androgenetic haploid embryonic stem cells

ABSTRACT: Epigenetic integrity of paternal imprints enhances the developmental potential of androgenetic haploid embryonic stem cells

PROVIDER: PRJNA707435 | ENA |

REPOSITORIES: ENA

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Epigenetic integrity of paternal imprints enhances the developmental potential of androgenetic haploid embryonic stem cells

Project description:The use of two inhibitors of Mek1/2 and Gsk3β (2i) promotes the generation of mouse diploid and haploid embryonic stem cells (ESCs) from the inner cell mass of biparental and uniparental blastocysts, respectively. However, a system enabling long-term maintenance of imprints in ESCs has proven challenging. Here, we report that usage of a two-step a2i (alternative two inhibitors of Src and Gsk3β, TSa2i) derivation/culture protocol results in the establishment of androgenetic haploid ESCs (AG-haESCs) with stable DNA methylation at paternal DMRs (differentially DNA methylated regions) up to passage 60 that can efficiently support generating mice upon oocyte injection. We also show coexistence of H3K9me3 marks and ZFP57 bindings with intact DMR methylations. Furthermore, we demonstrate that TSa2i-treated AG-haESCs are a heterogeneous cell population regarding paternal DMR methylation. Strikingly, AG-haESCs with late passages display increased paternal-DMR methylations and improved developmental potential compared to early-passage cells, in part through the enhanced proliferation of H19-DMR hypermethylated cells. Together, we establish AG-haESCs that can long-term maintain paternal imprints.

2022-01-26 | GSE168447 | GEO

Epigenetic integrity of paternal imprints enhances the developmental potential of androgenetic haploid embryonic stem cells.

Project description: Not available

| S-EPMC8783938 | biostudies-literature

CGH analysis of androgenetic haploid embryonic stem cells

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

High-frequency generation of fertile transgenic mice from H19-Igf2 engineered androgenetic haploid embryonic stem cells

Project description:Recent success in the derivation of mouse haploid embryonic stem cells from androgenetic blastocysts (ahESCs) has provided new avenues for the generation of genetically modified animals. However, the efficiency to produce viable transgenic mice via intracytoplasmic ahESCs injection (ICAI) was very low, which may correlate with the aberrant regulation of imprinted genes. Here we designed to delete the paternal imprinted gene H19 by CRSPR-Cas9 system combined with homologous recombination. The H19 deleted (H19Î) ahESCs maintained haploidy and genome integrity, expressed pluripotency markers, differentiated into embryoid bodies (EBs), and contributed to chimeras after blastocyst injection. These cells exhibited similar imprinting features with sperm cells, and can produce fertile progenies after ICAI at a high efficiency. More importantly, it is feasible to perform genetic manipulations in H19Î ahESCs, and the genomic modifications can be properly transmitted to offspring. Our study will benefit the reproductive medicine in curing the hereditary genetic diseases and infertility in the future. The copy number variations of the two H19Î ahESC lines were analyzed by the SurePrint G3 Mouse CGH 4Ã180 K microarrays (Agilent). Wild-type OG-3 ahESCs were used as reference.

2015-11-05 | E-GEOD-67563 | biostudies-arrayexpress

High-frequency generation of fertile transgenic mice from H19-Igf2 engineered androgenetic haploid embryonic stem cells

Project description:Recent success in the derivation of mouse haploid embryonic stem cells from androgenetic blastocysts (ahESCs) has provided new avenues for the generation of genetically modified animals. However, the efficiency to produce viable transgenic mice via intracytoplasmic ahESCs injection (ICAI) was very low, which may correlate with the aberrant regulation of imprinted genes. Here we designed to delete the paternal imprinted gene H19 by CRSPR-Cas9 system combined with homologous recombination. The H19 deleted (H19Δ) ahESCs maintained haploidy and genome integrity, expressed pluripotency markers, differentiated into embryoid bodies (EBs), and contributed to chimeras after blastocyst injection. These cells exhibited similar imprinting features with sperm cells, and can produce fertile progenies after ICAI at a high efficiency. More importantly, it is feasible to perform genetic manipulations in H19Δ ahESCs, and the genomic modifications can be properly transmitted to offspring. Our study will benefit the reproductive medicine in curing the hereditary genetic diseases and infertility in the future.

2015-11-05 | GSE67563 | 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. Genomic DNA of FACS sorted G0/G1 stage ahES cells was compared with 129/Sv male mouse kidney genomic DNA reference by the NimbleGen Mouse CGH 3x720K Whole-Genome Tiling Array (Build MM9) [100718_MM9_WG_CGH_HX3]

2012-08-30 | E-GEOD-39390 | biostudies-arrayexpress

CGH analysis of AG-haESCs (androgenetic haploid embryonic stem cells)

2012-04-27 | GSE35786 | GEO

CGH analysis of AG-haESCs (androgenetic haploid embryonic stem cells)

Project description:The genomic DNA sample of AG-haESCs were compared to the C57BL/6J male mouse kidney by comparative genomic hybridization. The data confirmed that the haploid cells sustained genome integrity. The analysis was performed on a NimbleGen Mouse CGH 3x720K Whole-Genome Tiling Array to analyse the copy number variations in AG-haESCs, and the genomic DNA of C57BL/6J male mouse kidney was used as control, which had the same background with haploid ESCs.

2012-04-26 | E-GEOD-35786 | biostudies-arrayexpress

Polycomb-mediated repression of paternal chromosomes maintains haploid dosage in diploid embryos of Marchantia [RNA-seq]

Project description:Complex mechanisms regulate gene dosage throughout eukaryotic life cycles. Mechanisms controlling gene dosage have been extensively studied in animals, however it is unknown how generalizable these mechanisms are to diverse eukaryotes. Here, we use the haploid plant Marchantia polymorpha to assess gene dosage control in its short-lived diploid embryo. We show that throughout embryogenesis, paternal chromosomes are repressed resulting in functional haploidy. The paternal genome is targeted for genomic imprinting by the Polycomb mark H3K27me3 starting at fertilization, rendering the maternal genome in control of embryogenesis. Maintaining haploid gene dosage by this new form of imprinting is essential for embryonic development. Our findings illustrate how haploid-dominant species can regulate gene dosage through paternal chromosome inactivation and initiates the exploration of the link between life cycle history and gene dosage in a broader range of organisms.

2022-08-23 | GSE193306 | GEO

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