Mus musculus strain:BDF1
Ontology highlight
ABSTRACT:
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| DRR045107.fastq.gz | Fastqsanger.gz | |||
| DRR045108.fastq.gz | Fastqsanger.gz | |||
| DRR045109.fastq.gz | Fastqsanger.gz | |||
| DRR045110.fastq.gz | Fastqsanger.gz | |||
| DRR045111.fastq.gz | Fastqsanger.gz |
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Project description:In mammals, many germline genes are repressed by epigenetic mechanisms to prevent their illegitimate expression in embryonic and somatic cells. To advance our understanding of the complete mechanisms restricting the expression of germline genes in mammals, we analyzed the chromatin signature of germline genes and performed a genome-wide CRISPR-Cas9 knock-out screen for genes involved in germline gene repression using a reporter system in which GFP is under the control of the epigenetically repressed Dazl germline promoter in mouse embryonic stem cells (ESCs). We showed that the repression of germline genes mainly depends on the polycomb complex PRC1.6 and DNA methylation, which function additively in mouse ESCs. Furthermore, we identified and validated several novel genes involved in the repression of germline genes, and characterized three of them: Usp7, Shfm1 (also known as Sem1) and Erh. Inactivation of Usp7, Shfm1 or Erh led to the upregulation of germline genes, as well as retrotransposons for Shfm1, in mouse ESCs. Functionally, Usp7 acts at two levels: firstly it associates with PRC1.6 components and represses germline genes independently of DNA methylation, and secondly it facilitates DNA methylation deposition at germline genes for long term repression. In summary, our study provides a global view of the epigenetic mechanisms and novel factors required for silencing germline genes in embryonic cells.
2023-02-03 | PXD030666 | Pride
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
Project description:DNA methylation erasure is required for mammalian primordial germ cell reprogramming. TET enzymes iteratively oxidize 5-methylcytosine to generate 5-hyroxymethylcytosine (5hmC), 5-formylcytosine, and 5-carboxycytosine to facilitate active genome demethylation. Whether these bases are required to promote replication-coupled dilution or activate base excision repair during germline reprogramming remains unresolved due to the lack of genetic models that decouple TET1 activities. Here, we generated two mouse lines expressing catalytically inactive TET1 (Tet1-HxD) and TET1 that stalls at 5hmC (Tet1-V). This experiment was done to conduct genome wide DNA methylation profiling of catalytically inactive Tet1HxD/HxD sperm, 5hmC stalling Tet1V/V sperm, KO Tet1-/- sperm, and WT sperm. Sperm samples were collected from adult males (>10 week).
2024-02-27 | GSE249772 | GEO
Project description:DNA methylation erasure is required for mammalian primordial germ cell reprogramming. TET enzymes iteratively oxidize 5-methylcytosine to generate 5-hyroxymethylcytosine (5hmC), 5-formylcytosine, and 5-carboxycytosine to facilitate active genome demethylation. Whether these bases are required to promote replication-coupled dilution or activate base excision repair during germline reprogramming remains unresolved due to the lack of genetic models that decouple TET1 activities. Here, we generated two mouse lines expressing catalytically inactive TET1 (Tet1-HxD) and TET1 that stalls at 5hmC (Tet1-V). This experiment was done to conduct genome wide DNA methylation profiling of catalytically inactive Tet1HxD/HxD sperm, 5hmC stalling Tet1V/V sperm, KO Tet1-/- sperm, and WT sperm. Sperm samples were collected from adult males (>10 week).
2024-02-27 | GSE224349 | GEO
Project description:Testicular toxicity is one of the frequent adverse effects of cancer chemotherapy and the problem is that there is no effective biomarker. To find effective biomarkers, we focused on epigenetic mechanisms of male germline. Therefore, our study investigated the DNA methylation status of male germline under the testicular toxicity induced by doxorubicin(DXR), a widely used anticancer agent. We established mouse models of initial stage of testicular toxicity and testicular pre-toxicity by administrating 0.2 mg/kg and 0.02 mg/kg of DXR twice a week for 5 weeks. Western blotting analysis revealed the protein expression levels of DNA methyltransferases DNMT3a and DNMT3b were decreased in both the DXR administration groups. Consistently, comprehensive DNA methylation analysis of sperm DNA using MBD-seq revealed that the majority of methylation changes induced by DXR administration were hypomethylation. This study showed the possibility of early diagnosis of testicular toxicity by examining DNA methylation status of sperm.
2019-02-04 | GSE110227 | GEO
Project description:In the male mouse germline, PIWI-interacting RNAs (piRNAs), bound by the PIWI protein MIWI2 (PIWIL4), guide DNA methylation of young active transposons through SPOCD1. However, the underlying mechanisms of SPOCD1-mediated piRNA-directed transposon methylation and whether this pathway functions to protect the human germline remains unknown. We identified loss-of-function variants in human SPOCD1 that cause defective transposon silencing and male infertility. Through the analysis of one of these pathogenic alleles, we discovered that the uncharacterised protein C19ORF84 interacts with SPOCD1. DNMT3C, the DNA methyltransferase responsible for transposon methylation, associates with SPOCD1 and C19ORF84 in foetal gonocytes. Furthermore, C19ORF84 is essential for piRNA-directed DNA methylation and male mouse fertility. Finally, C19ORF84 mediates the in vivo association of SPOCD1 with the de novo methylation machinery. In summary, we have discovered a conserved role for the human piRNA pathway in transposon silencing and C19ORF84, an uncharacterised protein essential for orchestrating piRNA-directed DNA methylation.
2024-02-14 | PXD047331 | Pride
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 | GSE292523 | GEO
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 | GSE293292 | GEO
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
Project description:DNA methylation erasure is required for mammalian primordial germ cell reprogramming. TET enzymes iteratively oxidize 5-methylcytosine to generate 5-hyroxymethylcytosine (5hmC), 5-formylcytosine, and 5-carboxycytosine to facilitate active genome demethylation. Whether these bases are required to promote replication-coupled dilution or activate base excision repair during germline reprogramming remains unresolved due to the lack of genetic models that decouple TET1 activities. Here, we generated two mouse lines expressing catalytically inactive TET1 (Tet1-HxD) and TET1 that stalls at 5hmC (Tet1-V). This experiment was done to conduct genome wide DNA methylation profiling of catalytically inactive Tet1HxD/HxD sperm, 5hmC stalling Tet1V/V sperm, KO Tet1-/- sperm, and WT sperm. Sperm samples were collected from adult males (>10 week). E17.5 WT prospermatogonia were collected using FACS of Oct4-GFP cells and CUT&RUN for H3K4me3 and IgG were done on 130,000 cells.
2024-02-27 | GSE224458 | GEO