Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

Genomic instability during reprogramming by nuclear transfer is DNA replication dependent

ABSTRACT: 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.

ORGANISM(S): Homo sapiens

PROVIDER: GSE93123 | GEO | 2017/01/05

SECONDARY ACCESSION(S): PRJNA360093

REPOSITORIES: GEO

ACCESS DATA

Json Xml

Similar Datasets

First round of DNA replication is essential for transcriptional reprogramming of somatic nuclei in mouse oocytes

Project description:Many of the structural and mechanistic requirements of oocyte-mediated nuclear reprogramming remain elusive. Previous accounts that transcriptional reprogramming of somatic nuclei in mouse zygotes may be complete in 24-36 hours, far more rapidly than in other reprogramming systems, raise the question of whether the mere exposure to the activated mouse ooplasm is sufficient to enact reprogramming in a nucleus. We therefore prevented DNA replication and cytokinesis, which ensue after nuclear transfer, in order to assess their requirement for transcriptional reprogramming of the key pluripotency genes Oct4 (Pou5f1) and Nanog in cloned mouse embryos. Using transcriptome and allele-specific analysis, we observed that hundreds of mRNAs, but not Oct4 and Nanog, became elevated in nucleus-transplanted oocytes without DNA replication. Progression through the first round of DNA replication was essential but not sufficient for transcriptional reprogramming of Oct4 and Nanog, whereas cytokinesis and thereby cell-cell interactions were dispensable for transcriptional reprogramming. Responses similar to clones also were observed in embryos produced by fertilization in vitro. Our results link the occurrence of reprogramming to a previously unappreciated requirement of oocyte-mediated nuclear reprogramming, namely DNA replication. Nuclear transfer alone affords no immediate transition from a somatic to a pluripotent gene expression pattern unless DNA replication is also in place. This study is therefore a resource to appreciate that the quest for always faster reprogramming methods may collide with a limit that is dictated by the cell cycle.

2016-12-17 | GSE53497 | GEO

Homo sapiens

Project description:Genomic instability during reprogramming by nuclear transfer is DNA replication dependent

| PRJNA360093 | ENA

Impeding Xist expression from the active X chromosome improves mouse somatic cell cloning

Project description:Cloning mammals by somatic cell nuclear transfer (SCNT) is highly inefficient because of aberrant genomic reprogramming. In addition to random reprogramming errors, we hypothesized the presence of specific errors as evidenced by common anomalies among clones. We found that Xist, which normally inactivates one of the two X chromosomes in females, was ectopically expressed from the active X (Xa) chromosome in cloned mouse embryos of both sexes. Deletion of Xist on Xa normalized global gene expression and produced about a 10-fold increase in cloning efficiency. We also identified an Xist-independent mechanism that specifically downregulated a subset of X-linked genes through somatic-type repressive histone blocks. Thus, we have identified nonrandom reprogramming errors in mouse cloning, which provide promising targets for breakthroughs in SCNT cloning technology. Gene expression were measured in mouse in vitro fertilized and somatic cell cloned blastocysts. More than three biological replicates were performed in each group using defferent nuclear donor cells.

2010-09-16 | E-GEOD-23181 | biostudies-arrayexpress

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

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

| PRJNA434693 | ENA

Impeding Xist expression from the active X chromosome improves mouse somatic cell cloning

2010-09-16 | GSE23181 | GEO

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

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

Global gene expression profiles of NT, AI and IVF bovine blastocysts, examination of nuclear reprogramming

Project description:Nuclear transfer (NT) has potential applications in agriculture and biomedicine, but the technology is hindered by low efficiency. Global gene expression analysis of clones is important for the comprehensive study of nuclear reprogramming. Here, we compared global gene expression profiles of individual bovine NT blastocysts with their somatic donor cells and fertilized control embryos utilizing cDNA microarray technology. The NT embryos’ gene expression profiles were drastically different from those of their donor cells and closely resembled those of the naturally fertilized embryos. Our findings demonstrate that the NT embryos have undergone significant nuclear reprogramming by the blastocyst stage; however, problems may occur during re-differentiation for tissue- and organogenesis, and small reprogramming errors may be magnified downstream in development. Keywords: cDNA microarray

2005-11-30 | GSE3568 | GEO

Mus musculus

Project description:Reprogramming is achieved within a single cell cycle after mouse nuclear transfer

| PRJNA138795 | ENA

Protein kinase C activation drives a differentiation program in an oligodendroglial precursor model through the modulation of specific biological networks

Project description:Protein kinase C (PKC) activation induces cellular reprogramming and differentiation in a variety of cellular models. Although many effectors of PKC physiological actions have been elucidated, the molecular mechanisms that regulate oligodendrocytes differentiation after PKC activation are still unclear. Here we applied a liquid chromatography - mass spectrometry (LC-MS/MS) approach to identify proteins, and biological processes, modulated after PKC activation in the established PKC-induced differentiation model, MO3.13 oligodendroglial cell line. Our analysis revealed that PKC signaling activation leads to the down-regulation of biological processes related to cell cycle and DNA replication and the up-regulation of a glial like developmental program. Our data suggested a role for ROCK in the modulation of MO3.13 phenotype, cytoskeletal and mechanotransduction dynamics but not in the modulation of cell cycle and calcium transport. Overall, our findings provides a resource for elucidating the PKC-mediated signaling network contributing to a more robust knowledge of the molecular dynamics leading to this cell fate transition.

2021-09-09 | PXD023336 | Pride

OmicsDI is part of the ELIXIR infrastructure

OmicsDI is an Elixir interoperability service. Learn more ›

OmicsDI Databases

PRIDE
PeptideAtlas
MassIVE
JPOST Repository
Physiome Model Repository

EGA
EVA
ENA
LINCS
PAXDB
Cell Collective

MetaboLights
Metabolomics Workbench
MetabolomeExpress
GNPS
BioModels
FAIRDOMHub

ArrayExpress
dbGaP
ExpressionAtlas
GEO
NODE

Information

Databases
Help
API
Contact us
Code on GitHub
Terms of use
Submit Data