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DNA methylation abnormal accumulation during cloned TSCs maintaining [RNA-seq]

ABSTRACT: Trophoblast stem cells (TSCs) are derived from the trophoectoderm of blastocysts and maintain ability to self-renewal and differentiation. TSC is a good model to research placenta development in vitro. It will contribute to understanding and improving cloned placentomegaly to compare the transcription and methylation between cloned and natural fertilized embryos throughout TSC derivation process. In the present study, we used SCNT (NT) and SCNT with HDACi treatment (SNT) as cloned groups and natural fertilized (NF) embryos to derive TSCs and chosed 5-time points to peform RNA-seq and RRBS. We found only NT got a barriar in TSC maintenace and both cloned groups exhited abnormal accumulating DNA methylation and it might be resiponsible for some malformations of cloned placentas.

ORGANISM(S): Mus musculus

PROVIDER: GSE163379 | GEO | 2021/03/01

REPOSITORIES: GEO

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DNA methylation abnormal accumulation during cloned TSCs maintaining [RRBS]

Project description:Trophoblast stem cells (TSCs) are derived from the trophoectoderm of blastocysts and maintain ability to self-renewal and differentiation. TSC is a good model to research placenta development in vitro. It will contribute to understanding and improving cloned placentomegaly to compare the transcription and methylation between cloned and natural fertilized embryos throughout TSC derivation process. In the present study, we used SCNT (NT) and SCNT with HDACi treatment (SNT) as cloned groups and natural fertilized (NF) embryos to derive TSCs and chosed 5-time points to peform RNA-seq and RRBS. We found only NT got a barriar in TSC maintenace and both cloned groups exhited abnormal accumulating DNA methylation and it might be resiponsible for some malformations of cloned placentas.

2021-03-01 | GSE163380 | GEO

Large-scale H3.1/H3.2 - H3K9me3 deposition defines the epigenetic signature of extraembryonic lineage cells [Repli-seq]

Project description:The extraembryonic lineage in mammals is unique in its unipotent differentiation ability and gestation-limited development. Here we identified the epigenomic characters of mouse trophoblast stem cells (TSCs) focusing on the genome-wide enrichment of histone H3.1/H3.2 and H3K9me3. Comparative ChIP-sequencing (ChIP-seq) analysis of TSCs and embryonic stem cells (ESCs) revealed that the TSC genome uniquely contained large (over 1Mb) H3.1/H3.2-H3K9me3 domains in the intergenic regions. This feature was common to extraembryonic cells in mice and humans. Depletion of CAF1, a H3.1/H3.2 chaperone, led to downregulation of TSC marker genes such as Cdx2 and Elf5 and ectopic expression of Oct3/4, an ESC marker. Nuclear transfer cloning using TSCs resulted in extremely poor embryonic development, but removal of H3K9me3 from their genome resulted in birth of the first TSC-cloned mice. Thus, the signature of the extraembryonic epigenome is safeguarded by the H3.1/H3.2-H3K9me3 enrichment, which protects the genome from cell fate transition.

2022-03-15 | GSE175935 | GEO

Large-scale H3.1/H3.2 - H3K9me3 deposition defines the epigenetic signature of extraembryonic lineage cells [HiC]

2022-03-15 | GSE175934 | GEO

Large-scale H3.1/H3.2 - H3K9me3 deposition defines the epigenetic signature of extraembryonic lineage cells [RNA-seq]

2022-03-15 | GSE175644 | GEO

Large-scale H3.1/H3.2 - H3K9me3 deposition defines the epigenetic signature of extraembryonic lineage cells [ChIP-seq]

2022-03-15 | GSE175638 | GEO

Large-scale H3.1/H3.2 - H3K9me3 deposition defines the epigenetic signature of extraembryonic lineage cells [ATAC-seq]

2022-03-15 | GSE175632 | GEO

Cellular dynamics of mouse trophoblast stem cells: Identification of a persistent stem cell type.

Project description:Mouse trophoblast stem cells (TSCs) proliferate indefinitely in vitro, despite their highly heterogeneous nature. In this study, we sought to characterize TSC colony types by using methods based on cell biology and biochemistry for a better understanding of how TSCs are maintained over multiple passages. Colonies of TSCs could be classified into four major types: type 1 is compact and dome-shaped, type 4 is flattened but with a large multilayered cell cluster, and types 2 and 3 are their intermediates. A time-lapse analysis indicated that type 1 colonies predominantly appeared after passaging, and a single type 1 colony gave rise to all other types. These colony transitions were irreversible, but at least some type 1 colonies persisted throughout culture. The typical cells comprising type 1 colonies were small and highly motile, and aggregated together to form primary colonies. A hierarchical clustering based on global gene expression profiles suggested that a TSC line containing more type 1 colony cells was similar to in-vivo extraembryonic tissues. Among the known TSC genes examined, Elf5 showed a differential expression pattern according to colony type, indicating that this gene might be a reliable marker of undifferentiated TSCs. When aggregated with fertilized embryos, cells from types 1 and 2, but not from type 4, distributed to the polar trophectoderm in blastocysts. These findings indicate that cells typically found in type 1 colonies can persist indefinitely as stem cells and are responsible for the maintenance of TSC lines. They may provide key information for future improvements in the quality of TSC lines.

2016-05-01 | GSE76255 | GEO

Transcription profiling of mouse 1 and 2 cell cloned embryos

Project description:Cloned embryos produced by somatic cell nuclear transfer (SCNT) display a plethora of phenotypic characteristics that make them different from fertilized embryos, indicating defects in the process of nuclear reprogramming by the recipient ooplasm. To elucidate the extent and timing of nuclear reprogramming, we used microarrays to analyze the transcriptome of mouse SCNT embryos during the first two cell cycles. We identified a large number of genes mis-expressed in SCNT embryos. We found that genes involved in transcription and regulation of transcription are prominent among affected genes, and thus may be particularly difficult to reprogram, and these likely cause a ripple effect that alters the transcriptome of many other functions, including oxidative phosphorylation, transport across membrane, and mRNA transport and processing. Interestingly, we also uncovered widespread alterations in the maternal (i.e. non transcribed) mRNA population of SCNT embryos. We conclude that gene expression in early SCNT embryos is grossly abnormal, and that this is at least in part the result of incomplete reprogramming of transcription factor genes. Experiment Overall Design: for both 1 and 2 cell stage we analyzed 4 samples each of fertilized embryos cultured with and without a-amanitin, scnt embryos cultured with and without a-amanitin, and parthennogenic embryos.

2008-06-15 | E-GEOD-6595 | biostudies-arrayexpress

RNA sequencing revealed abnormal gene expression and benefits of vitamin C treatment in cloned bovine embryos

Project description:Somatic cell nuclear transfer (SCNT) in mammals has been mostly unsuccessful. In this study, we measured and compared transcription profiles of cloned and fertilized embryos using RNA sequencing. Before zygotic genome activation (ZGA), fewer genes were activated in the cloned embryos than in vitro fertilization (IVF), leading to insufficient activation of protein transport and degradation functions. In blastocysts, terms of “embryo development” and “adherens junction” represented major differences for repressed genes in SCNT compared to IVF. Vitamin C (Vc) was found to relax donor cell chromatin and promote cloned embryo development. RNA sequencing indicated that Vc treatment restored some abnormal genes and processes in the cloned embryos. Processes of autophagy, RNA-editing and cell junction were not fully established, but they were improved by Vc treatment. Overexpression of ZNF641, one of the completely restored genes by Vc treatment, improved the cloned embryos’ potential. Finally, Vc treatment rescued some long non-coding RNAs that were aberrantly expressed in the cloned embryos. Collectively, many important genes and biological processes were abnormal in the cloned embryos, some of which can be improved by Vc treatment. This investigation provides several practical aspects that could be useful for improving cloning efficiency and its future applications.

2020-12-12 | GSE123705 | GEO

Gene expression profiling reveals a novel regulatory role for Sox21 in mouse trophoblast stem cell differentiation.

Project description:In this study we introduced Sox21 as a new regulator of trophoblast stem cell (TSC) differentiation. The transcriptome of different cell types were analyzed and compared to identify a TSC gene signature. In order to identify novel TSC specific genes, we performed genome-wide expression profiling of TSCs, embryonic stem cells, epiblast stem cells, and mouse embryo fibroblasts derived from mice of the same genetic background (129S1/SvImJ).

2015-11-10 | E-GEOD-67037 | biostudies-arrayexpress

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