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Reprogramming of chromatin accessibility in somatic cell nuclear transfer is DNA replication independent

ABSTRACT: Reprogramming of chromatin accessibility in somatic cell nuclear transfer is DNA replication independent

PROVIDER: PRJNA434693 | ENA |

REPOSITORIES: ENA

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Reprogramming of chromatin accessibility in somatic cell nuclear transfer is DNA replication independent

Project description:Mammalian oocytes have the ability to reset the transcriptional program of differentiated somatic cells into that of totipotent embryos through somatic cell nuclear transfer (SCNT). However, the mechanisms underlying SCNT-mediated reprogramming are largely unknown. To understand the mechanisms governing chromatin reprogramming during SCNT, we profiled DNaseI hypersensitive sites (DHSs) in donor cumulus cells and 1-cell stage SCNT embryos. To our surprise, the chromatin accessibility landscape of the donor cells is drastically changed to recapitulate that of the in vitro fertilization (IVF)-derived zygotes within 12 hours. Interestingly, this DHS reprogramming takes place even in the presence of a DNA replication inhibitor, suggesting that SCNT-mediated DHS reprogramming is independent of DNA replication. Thus, the study not only reveals the rapid and drastic nature of the changes in chromatin accessibility through SCNT, but also provides a DNA replication-independent model for studying cellular reprogramming.

2018-05-14 | GSE110851 | GEO

Reprogramming of Chromatin Accessibility in Somatic Cell Nuclear Transfer Is DNA Replication Independent.

Project description: Not available

| S-EPMC5988247 | biostudies-literature

Genomic instability during reprogramming by nuclear transfer is DNA replication dependent

Project description:Somatic cells can be reprogrammed to a pluripotent state by nuclear transfer into oocytes, yet developmental arrest often occurs. While incomplete transcriptional reprogramming is known to cause developmental failure, reprogramming also involves concurrent changes in gene expression, cell cycle progression and nuclear structure. Here we study cellular reprogramming events in human and mouse nuclear transfer (NT) embryos prior to embryonic genome activation. We show that genetic instability marked by frequent chromosome segregation errors and DNA damage arise prior to, and independent of, transcriptional activity. These errors occur following transition through DNA replication and are repaired by BRCA1. In the absence of mitotic nuclear remodeling, DNA replication is delayed and errors are exacerbated in subsequent mitosis. These results demonstrate that independent of gene expression, cell type-specific features of cell cycle progression constitute a barrier sufficient to prevent the transition from one cell type to another during reprogramming.

2017-01-05 | GSE93123 | GEO

Expression data from somatic cell nuclear transfer bovine embryo

Project description:Expression data from somatic cell nuclear transfer bovine embryo

| PRJNA351837 | ENA

Chromatin architecture reorganization in somatic cell nuclear transfer embryos

Project description:Chromatin architecture reorganization in somatic cell nuclear transfer embryos

| PRJNA608976 | ENA

Human embryonic stem cells derived by somatic cell nuclear transfer

Project description:In this study, we derived human embryonic stem cell via SCNT. Transcriptional analysis was performed to compare characteristics of SCNT-derived ESCs to their IVF counterpart. The transcriptomes of human embryonic stem cells (hESO-NT1) derived from human skin fibroblasts (HDF-f) via somatic cell nuclear transfer (SCNT) was compared with IVF-derived counterpart (hESO-7) derived using oocytes from identical donor and parental skin fibroblast (HDF-f). . Three biological replicates of each cell line (A, B, C) were analyzed.

2013-05-15 | E-GEOD-46397 | biostudies-arrayexpress

Genome-wide comparison of EpiSC lines derived from fertilized (FT) and somatic cell nuclear transfer (NT) embryos

Project description:Genome wide comparison of gene expression between EpiSC lines derived from fertilized (FT) embryos and somatic cell nuclear transfer (NT) embryos. EpiSC lines were derived from fertilized and somatic cell nuclear transfer embryos and cultured until 15 to 20 passages. RNA was then extracted in order to compare transcriptomic profiles.

2010-02-05 | GSE17402 | GEO

KMT1A (SUV39H1) prevents somatic cell nuclear transfer (SCNT) mediated reprogramming

Project description:KMT1A (also known as SUV39H1)was involved in improving reprogramming efficiency during inducing pluripotent stem cells (iPSC). However, the knowledge about KMT1A regulating somatic cell nuclear transfer (SCNT) mediated reprogramming was limited. To understand the role of KMT1A during SCNT embryos in vitro development; we performed based transcriptome profiling in SCNT embryos at 8-cell stages and KMT1A knockdown 8-cell SCNT embryos. The data show the mRNAs regulated after KMT1A was down-regulated.

2017-07-31 | GSE99072 | GEO

Histone Demethylase-Assisted Somatic Cell Nuclear Transfer Facilitates Derivation of Human Pluripotent Stem Cells

Project description:The extremely low efficiency of human embryonic stem cell (hESC) derivation using somatic cell nuclear transfer (SCNT) limits potential application. Blastocyst formation from human SCNT embryos occurs at a low rate and with only some oocyte donors. We previously showed in mice that reduction of histone H3 lysine 9 trimethylation (H3K9me3) through ectopic expression of the H3K9me3 demethylase Kdm4d greatly improves SCNT embryo development. Here we show that overexpression of a related H3K9me3 demethylase KDM4A improves human SCNT, and that, as in mice, H3K9me3 in the human somatic cell genome is an SCNT reprogramming barrier. Overexpression of KDM4A significantly improves the blastocyst formation rate in human SCNT embryos by facilitating transcriptional reprogramming, allowing derivation of NTESCs from all oocyte donors tested using adult AMD patient somatic nuclei donors. This conserved mechanistic insight has potential applications for improving SCNT in a variety of contexts, including regenerative medicine. Here we perform RNA-seq based transcriptome profiling in human Donor (fibroblast cells), in vitro fertilized embryos at 8-cell stages (IVF_8Cell), somatic cell nuclear transfer embryos at 8-cell stages (SCNT_8Cell), SCNT assisted by KDM4A 8-cell embryos (SCNT_KDM4A_8Cell). Besides, we also perform RNA-seq in Control human ES cells (CTR_hES) and SCNT assisted by KDM4A derived human ES cells (NTK) with duplicates.Â

2016-03-01 | E-GEOD-73362 | biostudies-arrayexpress

Reprogramming of fish somatic cells for nuclear transfer is primed by Xenopus egg extract

Project description:Somatic cell reprogramming in vitro prior to nuclear transfer is one strategy expected to improve clone survival during development. In this study, we investigated the reprogramming extent of fish fin somatic cells after in vitro exposure to Xenopus egg extract and subsequent culture. Using a cDNA microarray approach, we observed drastic changes in the gene expression profile of the treated cells. Several actors of the TGFbeta and Wnt/beta-catenin signaling pathways, as well as some mesenchymal markers, were inhibited in treated cells, while several epithelial markers were upregulated. This was associated with morphological changes of the cells in culture, suggesting that egg extract drove somatic cells towards a mesenchymal-epithelial transition (MET), the hallmark of somatic reprogramming in iPSCs. However, treated cells were also characterized by a strong decrease in de novo lipid biosynthesis metabolism, the lack of re-expression of pou2 and nanog pluripotency markers, and absence of DNA methylation remodeling of their promoter region. In all, this study showed that Xenopus egg extract treatment initiated an in vitro reprogramming of fin somatic cells in culture. Although not thorough, the induced changes have primed the somatic chromatin for a better embryonic reprogramming upon nuclear transfer.

2023-01-01 | GSE205854 | GEO

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