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CRISPR-Cas9 Screens Reveal Genes Regulating a G0-like State in Human Neural Progenitors [Bulk RNA-seq]

ABSTRACT: CRISPR-Cas9 Screens Reveal Genes Regulating a G0-like State in Human Neural Progenitors [Bulk RNA-seq]

PROVIDER: PRJNA480709 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR7511940_1.fastq.gz	Fastqsanger.gz
	SRR7511940_2.fastq.gz	Fastqsanger.gz
	SRR7511950_1.fastq.gz	Fastqsanger.gz
	SRR7511950_2.fastq.gz	Fastqsanger.gz
	SRR7511960_1.fastq.gz	Fastqsanger.gz

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CRISPR-Cas9 Screens Reveal Genes Regulating a G0-like State in Human Neural Progenitors [CRISPR]

Project description:The coordination of developmental potential and proliferation in stem and progenitor cells is essential for mammalian development and tissue homeostasis. To better understand this coordination in human neural progenitor cells (hNPCs), we performed CRISPR-Cas9 screens and identified genes that limit their expansion. These screens revealed that knockout of growth-limiting genes, including CREBBP, NF2, PTPN14, TAOK1, or TP53, caused increased hNPC expansion via skipping of a transient G0-like state, accompanied by transcriptional reprogramming of G1 subpopulations. Hallmarks of the G0-like state included expression of genes associated with quiescent neural stem cells and neural development and molecular features found in quiescent cells (e.g., hypo-phosphorylated Rb, CDK2low activity, and p27high). Further, G0-skip genes act through both distinct and convergent downstream effectors, including cell cycle, Hippo-YAP, and novel targets. The results suggest that hNPC expansion is constrained by a transient G0-like state, regulated by multiple pathways, that facilitates retention of neurodevelopmental identity.

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CRISPR-Cas9 Screens Reveal Genes Regulating a G0-like State in Human Neural Progenitors

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CRISPR-Cas9 Screens Reveal Genes Regulating a G0-like State in Human Neural Progenitors [CRISPR]

Project description:CRISPR-Cas9 Screens Reveal Genes Regulating a G0-like State in Human Neural Progenitors [CRISPR]

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CRISPR-Cas9 Screens Reveal Genes Regulating a G0-like State in Human Neural Progenitors [scRNA-seq]

Project description:CRISPR-Cas9 Screens Reveal Genes Regulating a G0-like State in Human Neural Progenitors [scRNA-seq]

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Project description:To identify new therapeutic targets for Glioblastoma (GBM), we performed genome-wide CRISPR-Cas9 "knockout" (KO) screens in patient-derived GBM stem-like cells (GSCs) and human neural stem/progenitors (NSCs), non-neoplastic stem cell controls, for genes required for their in vitro growth. Surprisingly, the vast majority GSC-lethal hits were found outside of molecular networks commonly altered in GBM and GSCs (e.g., oncogenic drivers). In vitro and in vivo validation of GSC-specific targets revealed several strong hits, including the wee1-like kinase, PKMYT1/Myt1. Mechanistic studies demonstrated that PKMYT1 acts redundantly with WEE1 to inhibit Cyclin B-CDK1 activity via CDK1-Tyr15 phosphorylation and to promote timely completion of mitosis in NSCs. However, in GSCs, this redundancy is lost, likely as a result of oncogenic signaling, causing GBM-specific lethality. A whole-genome CRISPR-Cas9 knockout screens targeting over 18,000 genes using the all-in-one LV-sgRNA:Cas9 platform system were performed using a âshot gunâ approach by transducing 2 GBM patient-derived isolates and 2 human neural stem cell isolates with the pool library (2 biological replicates), and cultures were outgrown for ~3 weeks. The end time point of each screen was compared to day 0 in order to determine which sgRNAs were overrepresented or underrepresented in the population.

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CRISPR-Cas9 Screens Reveal Genes Regulating a G0-like State in Human Neural Progenitors

Project description:This SuperSeries is composed of the SubSeries listed below.

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