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Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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RNA-seq of control siRNA, p150 siRNA and p60 siRNA transfected mES cells

ABSTRACT: Totipotent cells that resemble 2-cell stage embryos (2C-like cells) can arise with a very low frequency in ES cell cultures. Here we show that totipotent-like cells can be induced in vitro through the down-regulation of the chromatin assembly activity of CAF-1. Endogenous retroviruses and 2-cell stage specific genes are among the highest upregulated genes upon CAF-1 knockdown and the emerging totipotent-like cells exhibit the molecular characteristics of endogenous 2C-like cells and 2-cell embryos. CAF-1 depletion leads to increased accessibility at MERVL elements and to the up-regulation of neighbouring genes, thereby generating a transcriptional profile similar to that of 2-cell stage embryos.

INSTRUMENT(S): Illumina HiSeq 2500

ORGANISM(S): Mus musculus

SUBMITTER: Juan Vaquerizas

PROVIDER: E-MTAB-2684 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Early embryonic-like cells are induced by downregulating replication-dependent chromatin assembly.

Ishiuchi Takashi T Enriquez-Gasca Rocio R Mizutani Eiji E Bošković Ana A Ziegler-Birling Celine C Rodriguez-Terrones Diego D Wakayama Teruhiko T Vaquerizas Juan M JM Torres-Padilla Maria-Elena ME

Nature structural & molecular biology 20150803 9

Cellular plasticity is essential for early embryonic cells. Unlike pluripotent cells, which form embryonic tissues, totipotent cells can generate a complete organism including embryonic and extraembryonic tissues. Cells resembling 2-cell-stage embryos (2C-like cells) arise at very low frequency in embryonic stem (ES) cell cultures. Although induced reprogramming to pluripotency is well established, totipotent cells remain poorly characterized, and whether reprogramming to totipotency is possible ...[more]

PMID: 26237512

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NR5A2 connects genome activation to the first lineage segregation in totipotent embryos

Project description:Zygotic genome activation (ZGA) marks the beginning of the embryonic program for a totipotent embryo, which gives rise to inner cell mass (ICM), where pluripotent epiblast arises, and extraembryonic trophectoderm. While much has been learned about pluripotency regulation, how ZGA is connected to the first lineage segregation in early embryos remains elusive. Here, we investigated the role of nuclear receptor transcription factors (NR TFs), whose motifs are highly enriched in accessible chromatin at the 2-cell (2C) to 8-cell (8C) stages after ZGA. We found NR5A2, an NR TF highly induced upon ZGA, is required for early development. Genome-wide chromatin binding in 2C and 8C embryos showed NR5A2 bound cis-regulatory elements which strongly enrich B1 elements where its motif is embedded. RNA-seq showed while the zygotic genome was largely activated at the 2C stage, 4-8C-specific gene activation was substantially impaired in Nr5a2 KD embryos. At the 8C stage, NR5A2 preferentially regulated its targets including a subset of early ICM genes. Consistently, NR5A2 was required for opening 8C-specific binding sites. Interestingly, while NR5A2’s 2C-binding sites are largely closed in blastocyst, 8C-specific binding sites tend to stay open and are bound by master pluripotency TFs (NANOG, SOX2, and OCT4) in blastocyst or ESC. Further analyses showed NR5A2 regulates early ICM genes at the 8C stage, master pluripotency TFs regulate both early and late ICM genes at later stages. Taken together, these data identify NR5A2 as a key regulator in totipotent embryos that connects ZGA to the first lineage segregation in early mouse development.

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Global analysis of expression upon RNA Pol I inhibition (iPol I) in ESCs

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A transcriptional roadmap for 2C-like to pluripotent state transition [bulk RNA]

Project description:In mouse embryonic stem cell (ESC) culture, a small proportion of cells display totipotent features by expressing a set of genes that are only active in 2-cell-stage embryos. These 2-cell-like (2C-like) cells spontaneously transit back into pluripotent state. We previously dissected the transcriptional dynamics of pluripotent to 2C-like transition and identified factors that modulate the transition. However, how 2C-like cells transits back to the pluripotent state and what factors drive this process remains largely unknown. To address these questions, we examined the transcriptional dynamics during the reverse transition from the 2C-like state to ESCs and identified an intermediate state involved in the transition. Interestingly, we found that mESCs exit from the 2C-like state through a molecular path characterized by a two-wave upregulation of pluripotent genes different from the one observed during the 2C-like entry transition. We also showed that nonsense-mediated mRNA decay (NMD) targets Dux mRNA and affects 2C-like state maintenance, suggesting that Dux degradation contributes to the reversal of 2C-like state.

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