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TP53 promotes lineage commitment of human embryonic stem cells through ciliogenesis and sonic hedgehog signaling [CRISPR screen]

ABSTRACT: CRISPR-Cas9 screen was done to identify genes required for NPC differentiation. Sox1 is an established NPC marker. Human ESCs were infected with GecKO v2 library, selected with puromycin and then allowed to differentiate into Neural progenitor cells (NPC) using StemDiff monolayer protocol (Stem cell Technologies). Day 7 NPCs were collected and FACS sorted to obtain bottom 1% of Sox1 expressing cells. NGS was used to identify guide sequences in cell population and determine the genes required for Sox1 expression during NPC differentiation.

ORGANISM(S): Homo sapiens

PROVIDER: GSE168585 | GEO | 2022/02/16

REPOSITORIES: GEO

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TP53 promotes lineage commitment of human embryonic stem cells through ciliogenesis and sonic hedgehog signaling.

Project description:Aneuploidy, defective differentiation, and inactivation of the tumor suppressor TP53 all occur frequently during tumorigenesis. Here, we probe the potential links among these cancer traits by inactivating TP53 in human embryonic stem cells (hESCs). TP53-/- hESCs exhibit increased proliferation rates, mitotic errors, and low-grade structural aneuploidy; produce poorly differentiated immature teratomas in mice; and fail to differentiate into neural progenitor cells (NPCs) in vitro. Genome-wide CRISPR screen reveals requirements of ciliogenesis and sonic hedgehog (Shh) pathways for hESC differentiation into NPCs. TP53 deletion causes abnormal ciliogenesis in neural rosettes. In addition to restraining cell proliferation through CDKN1A, TP53 activates the transcription of BBS9, which encodes a ciliogenesis regulator required for proper Shh signaling and NPC formation. This developmentally regulated transcriptional program of TP53 promotes ciliogenesis, restrains Shh signaling, and commits hESCs to neural lineages.

| S-EPMC8904926 | biostudies-literature

Mus musculus

Project description:Sox1 mouse NPC ChIP-seq

| PRJNA389178 | ENA

TP53 promotes lineage commitment of human embryonic stem cells through ciliogenesis and sonic hedgehog signaling

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

2022-02-15 | GSE168587 | GEO

TP53 promotes lineage commitment of human embryonic stem cells through ciliogenesis and sonic hedgehog signaling [RNA-Seq]

Project description:H9 WT ESCs or TP53 KO (Clone 1) ESCs were injected subcutaneously into mice for teratoma formation. 6-8 weeks later, teratomas were isolated and total RNA was purified. TruSeq library preparation kit from Illumina was used to make libraries for RNA-Seq. NGS sequencing on Illumina NextSeq 500 was performed and data was analyzed to determine changes in transcriptome between WT and TP53 KO teratoma cells.

2022-02-16 | GSE168678 | GEO

TP53 promotes lineage commitment of human embryonic stem cells through ciliogenesis and sonic hedgehog signaling [ChIP-seq]

Project description:NGS was used to analyze role of TP53 in differentiation of human embryonic stem cells. TP53, a transcription factor, binds DNA and regulates transcription of specific genes. Chipseq of TP53 was done on WT ESCs and NPCs to identify direct targets of TP53 during differentiation of hESCs into neuronal lineage.

2022-02-16 | GSE168584 | GEO

TP53 promotes lineage commitment of human embryonic stem cells through ciliogenesis and sonic hedgehog signaling [RNA-Seq 2]

Project description:NGS was used to analyze role of TP53 in differentiation of human embryonic stem cells. RNAseq and was done in WT and p53 KO embryonic stem cells (ESCs) and in in vitro differentiated Neural progenitor cells (NPCs).

2022-02-16 | GSE168752 | GEO

RNA-sequencing and CRISPR/Cas9 Screen of PEO1 cells grown in suspension.

Project description:RNA-sequencing of PEO1 cells grown in suspension (SUS1 and SUS3) was performed to evaluate transcriptional reprogramming during anokis escape. CRISPR/Cas9 screen of PEO1 cells grown in adherent and suspension culture settings for 5 days or 10 days. GeCKO v2 library transduced in PEO1 cell line and cells were selected with puromycin. PEO1 line was authenticated prior to sequencing. PEO1 parental were confirmed to be BRCA2-mutated (5139C>G).

2018-12-04 | GSE123290 | GEO

SOX1 Determines the Regional Identity of Neural Progenitors Differentiated from Human Embryonic Stem Cells

Project description:During human embryogenesis, primitive neural cells start to be generated at the time of gastrulation and gradually acquire regional identities, which is a process called neural patterning. But how intrinsic factors respond to exogenous patterning signals remains poorly understood. Human Embryonic Stem Cells (hESCs) provide a great model to recapitulate this process. Through exogenous manipulation of canonical WNT signaling activation during neural differentiation, dose-dependent specification of regionally defined neural progenitors ranging from the telencephalic forebrain to posterior hindbrain could be rapidly and efficiently induced. Unexpectedly, we find that SOX1, generally referred as a pan-neural gene, displays a regional specific distribution in the human neural patterning process. To investigate the expression and function of SOX1 efficiently, we have generated the SOX1-EGFP reporter and SOX1-knockout (KO) hESC lines using the CRISPR/Cas9 system. SOX1 is initially expressed across the mes–met border and peaked in the metencephalon region at the early regional specification stage. Its depletion leads to the posterior shift of the mes–met border. Therefore, SOX1 is required to define the border at a correct region. In-depth analysis of SOX1 ChIP-sequencing and transcriptome data will provide more insights into how SOX1 determines the mes-met border formation and identify the downstream targets of SOX1. This study identifies SOX1 as one of the intrinsic factors key for the prepattern establishment in the developing central nervous system, particularly for defining the isthmus position.

2020-08-26 | GSE138217 | GEO

Transcriptional profiling by 4SU-seq in mouse ESCs and ESC-derived neural progenitor cells.

Project description:Nascent RNA was metabolically labelled with 4SU in undifferentiated and ESC-derived neural progenitor cells (NPCs). 4SU incorporated RNA was isolated and deep-sequenced at day 0 (ESCs), 3, 5 and 7 of differentiation. NPC differentiation was monitored through expression of a GFP reporter insereted into the Sox1 locus (46C reporter ESC line; PMID: 12524553). The aim was to monitor changes in transcription as ESCs differentiate into NPCs and relate this to enhancer activity.

2019-01-12 | GSE115774 | GEO

Gene expression profiles of nasopharyngeal carcinoma cell line HNE2 following TP53 overexpression

Project description:Recent studies have revealed that long non-coding RNAs (lncRNAs) participate in all steps of cancer initiation and progression by regulating protein coding genes at the epigenetic, transcriptional and post-transcriptional levels. LncRNAs are in turn regulated by other genes, forming a complex regulatory network. The regulation networks between the p53 tumor suppressor and lncRNAs in nasopharyngeal carcinoma (NPC) remain unclear. The aim of this study was to investigate the regulatory roles of the TP53 gene in regulating lncRNA and mRNA expression profiles in NPC cell line HNE2. p53 induced gene expression in human nasopharyngeal carcinoma cell line HNE2 was measured at 0, 12, 24 and 48 hours after transfected by pCMV-p53 plasmid.

2015-05-14 | E-GEOD-60379 | biostudies-arrayexpress

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