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Tracing the transitions from pluripotency to germ cell fate with CRISPR screening

ABSTRACT: Tracing the transitions from pluripotency to germ cell fate with CRISPR screening

PROVIDER: PRJNA482361 | ENA |

REPOSITORIES: ENA

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Tracing the transitions from pluripotency to germ cell fate with CRISPR screening

Project description:Early mammalian development entails a series of cell fate transitions that includes transit through naïve pluripotency to post-implantation epiblast. This can subsequently give rise to primordial germ cells (PGC), the founding population of the germline lineage. To investigate the gene regulatory networks that control these critical cell fate decisions, we developed a compound-reporter system to track cellular identity in a model of PGC specification (PGC-like cells; PGCLC), and coupled it with unbiased genome-wide CRISPR screening. This enabled identification of key genes both for exit from pluripotency and for acquisition of PGC fate, with further characterisation revealing a central role for the transcription regulators Nr5a2 and Zfp296 in germline ontogeny. Abrogation of these genes results in significantly impaired PGCLC development accompanied with widespread activation (Nr5a2-/-) or inhibition (Zfp296-/-) of WNT pathway components. This leads to aberrant upregulation of the somatic programme or failure to appropriately activate germline genes in PGCLC, respectively, and consequently loss of germ cell identity. Overall our study places Zfp296 and Nr5a2 as key components of an expanded PGC gene regulatory network, and outlines a transferable strategy for identifying critical regulators of complex cell fate transitions.

2018-10-01 | GSE117473 | GEO

Tracing the transitions from pluripotency to germ cell fate with CRISPR screening.

Project description: Not available

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RNA Pol II pausing facilitates phased pluripotency transitions by buffering transcription

Project description:PRO-seq experiment of Nelfb deg mESC in serum/LIF or during pluripotency transitions, with or without dTAG-13.

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Tracing the conversion process for primordial germ cells to pluripotent stem cells

Project description:The overall gene expression profiles revealed by this system provide novel insight into how pluripotency is acquired in germ-committed cells. In the study presented here, we employed FACS sorting to purify pluripotent candidate cells during the culture period

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Tracing the conversion process for primordial germ cells to pluripotent stem cells

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The H3K27 demethylase Utx facilitates somatic and germ cell epigenetic reprogramming to pluripotency

Project description:This SuperSeries is composed of the following subset Series: GSE35775: The H3K27 demethylase Utx facilitates somatic and germ cell epigenetic reprogramming to pluripotency [Affymetrix gene expression] GSE37821: The H3K27 demethylase Utx facilitates somatic and germ cell epigenetic reprogramming to pluripotency [ChIP-Seq] Refer to individual Series

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Distinct mechanisms of germ cell factor regulation for an inductive germ cell fate

Project description:Here we employed single cell RNA sequencing to identify the transcriptional program of Nanos and Vasa positive cells and their changes during development. Our single cell sequencing analysis of six developmental stages in P. miniata revealed cell types derived from the three germ layers and expression of the germ cell genes Nanos and Vasa. We used these datasets to parse out 20 cell lineages of the embryo identified by this approach and to focus on the key transitions of germ cell gene expression and test their coexpression with key signaling components.

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Multi-Omic Profiling Reveals Dynamics of the Phased Progression of Pluripotency

Project description:Pluripotency is highly dynamic and progresses through a continuum of pluripotent stem-cell states. The two states that bookend the pluripotency continuum, naïve and primed, are well characterized, but our understanding of the intermediate states and transitions between them remain incomplete. Here, we dissect the dynamics of pluripotent state transitions underlying pre- to post-implantation epiblast differentiation. Through comprehensive mapping of the proteome, phosphoproteome, transcriptome, and epigenome of embryonic stem cells transitioning from naïve to primed pluripotency, we find that rapid, acute, and widespread changes to the phosphoproteome precede ordered changes to the epigenome, transcriptome, and proteome. Reconstruction of kinase-substrate networks reveals signaling cascades, dynamics, and crosstalk. Distinct waves of global proteomic changes mark discrete phases of pluripotency, with cell state-specific surface markers tracking pluripotent state transitions. Our data provide new insights into the multi-layered control of the phased progression of pluripotency and a foundation for modeling mechanisms regulating pluripotent state transitions (www.stemcellatlasorg).

2019-05-07 | PXD010621 | Pride

RNA Pol II pausing facilitates phased pluripotency transitions by buffering transcription

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