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A CRISPR/Cas9 screen in embryonic stem cells reveals that Mdm2 regulates totipotency exit [CUT&Tag]

ABSTRACT: A CRISPR/Cas9 screen in embryonic stem cells reveals that Mdm2 regulates totipotency exit [CUT&Tag]

PROVIDER: PRJNA1120447 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR29297846_1.fastq.gz	Fastqsanger.gz
	SRR29297846_2.fastq.gz	Fastqsanger.gz
	SRR29297847_1.fastq.gz	Fastqsanger.gz
	SRR29297847_2.fastq.gz	Fastqsanger.gz
	SRR29297848_1.fastq.gz	Fastqsanger.gz

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Similar Datasets

A CRISPR/Cas9 screen in embryonic stem cells reveals that Mdm2 regulates totipotency exit [CUT&Tag]

Project description:During early embryonic development, the transition from totipotency to pluripotency is a fundamental and critical process for proper development. However, the regulatory mechanisms governing this transition remain elusive. In this study, we conducted a comprehensive genome-wide CRISPR/Cas9 screen to investigate the 2-cell-like cells (2CLCs) phenotype in mouse embryonic stem cells (mESCs). This effort led to the identification of ten regulators that play a pivotal role in determining cell fate during this transition. Notably, our study revealed Mdm2 as a significant negative regulator of 2CLCs, as perturbation of Mdm2 resulted in a higher proportion of 2CLCs. Mdm2 appears to influence cell fate through its impact on cell cycle progression and H3K27me3 epigenetic modifications. In summary, the results of our CRISPR/Cas9 screen have uncovered several genes with distinct functions in regulating totipotency and pluripotency at various levels, offering a valuable resource for potential targets in future molecular studies.

2024-07-01 | GSE269169 | GEO

A CRISPR/Cas9 screen in embryonic stem cells reveals that Mdm2 regulates totipotency exit [RNA-Seq]

2024-07-01 | GSE269168 | GEO

A CRISPR/Cas9 screen in embryonic stem cells reveals that Mdm2 regulates totipotency exit [RNA-Seq]

Project description:A CRISPR/Cas9 screen in embryonic stem cells reveals that Mdm2 regulates totipotency exit [RNA-Seq]

| PRJNA1120448 | ENA

The homeobox transcription factor DUXBL controls exit from totipotency

Project description:The homeobox transcription factor DUXBL controls exit from totipotency

| PRJNA979244 | ENA

The homeobox transcription factor DUXBL controls exit from totipotency

Project description:In mice, exit from the totipotent 2-cell (2C) embryo stage requires silencing of the 2C-associated transcriptional program. However, the molecular mechanisms involved in this process remain poorly understood. Here, we demonstrate that the 2C-specific transcription factor DUX mediates an essential negative feedback loop by inducing the expression of DUXBL to promote this silencing.We show that DUXBL gains accessibility to DUX-bound regions specifically upon DUX expression.Furthermore, we determine that DUXBL interacts with TRIM24 and TRIM33, members of the tripartite motif superfamily involved in gene silencing and co-localizes with them in nuclear foci upon DUX expression. Importantly, DUXBL overexpression impairs 2C-associated transcription, whereas Duxbl inactivation in ESC increases DUX-dependent induction of the 2C-transcriptional program. Consequently, DUXBL deficiency in embryos results in sustained expression of 2C-associated transcripts, leading to early developmental arrest.Our study identifies DUXBL as an essential regulator of totipotency exit enabling the first divergence of cell fates.

2023-12-07 | GSE234053 | GEO

Zygotic genome activation by the totipotency pioneer factor Nr5a2 [CUT&Tag]

Project description:Zygotic genome activation by the totipotency pioneer factor Nr5a2 [CUT&Tag]

| PRJNA738283 | ENA

Defining totipotency using criteria of increasing stringency

Project description:Totipotency is the ability of a single cell to give rise to all the differentiated cells that build the conceptus, yet how to capture this property in vitro remains incompletely understood. Defining totipotency relies upon a variety of assays of variable stringency. Here we describe criteria to define totipotency. We illustrate how distinct criteria of increasing stringency can be used to judge totipotency by evaluating candidate totipotent cell types in the mouse, including early blastomeres and expanded or extended pluripotent stem cells. Our data challenge the notion that expanded or extended pluripotent states harbor increased totipotent potential relative to conventional embryonic stem cells under in vivo conditions.

2020-10-13 | GSE145609 | GEO

A satellite DNA array barcodes chromosome 7 and regulates totipotency via ZFP819

Project description:Mammalian genomes are a battleground for genetic conflict between repetitive elements (REs) and KRAB-zinc finger proteins (KZFPs). We explored whether KZFPs can regulate cell fate by employing Zfp819, which targets a satellite DNA array, ZP3AR. ZP3AR coats a megabase region of chromosome 7 that encompasses genes encoding ZSCAN4, a master transcription factor of totipotency. Depleting Zfp819 in mouse embryonic stem cells (mESCs) causes them to transition to a 2-cell (2C)-like state, whereby the ZP3AR array switches from a poised to an active enhancer state. This is accompanied by a global eviction of heterochromatin roadblocks, which we link to decreased SETDB1 stability. These events result in transcription of active LINE-1 elements and a block in differentiation. In sum, ZFP819 and TRIM28 partner up to close chromatin across Zscan4, allowing exit from totipotency. We propose that satellite DNAs may control developmental fate transitions through the barcoding and switching off of master transcription factor genes.

2022-10-31 | GSE197548 | GEO

Genome-wide maps of chromatin binding site for Mdm2.

Project description:The MDM2 oncoprotein is an essential component of the p53 pathway. Although MDM2 has been widely described as a major negative regulator of the p53 tumor suppressor, growing evidences support the notion that Mdm2 functions are also mediated through p53 independent mechanisms. We found that MDM2 is recruited on chromatin. By ChIP-Seq (using MDM2 antibody) of H1299 cells exhibiting MDM2 recruitment on chromatin, we identified at the whole genome level MDM2 binding sites. Both input and IPped-DNA were exhaustively sequenced and mapped on human genome

2016-06-09 | GSE64439 | GEO

Duxbl-mediated suppression of ZGA drives totipotency to pluripotency transition in mice III

Project description:In mammalian embryos, Dux transcription factors (TF) drive a cleavage stage-specific transcriptional burst associated with zygotic genome activation (ZGA) during which hundreds of genes and endogenous retroviral (ERV) elements are transiently expressed. In mice, ZGA begins in late 1-cell (1C) embryos and is shut down within hours at the late 2-cell (2C) to 4-cell (4C) stage. While this transition is accompanied by the loss of totipotency, it is not clear whether shutdown of Dux-induced ZGA is required for this process and how ZGA is terminated. Here, we reveal an essential negative feedback axis by which the Dux family member Duxbl terminates Dux-induced ZGA from the mid 2C stage onward by direct suppression of genomic Dux target loci. Consequently, Duxbl inactivation results in sustained expression of Dux-induced ZGA leading to 4C arrest. Our study reveals the Dux/Duxbl axis that determines the timing of totipotency exit enabling the very first divergence of cell fates.

2023-12-07 | GSE213702 | GEO

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