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Genome-wide identification of target genes for the histone demethylase Phf8 in the oligodendroglial cell line CG4 [Phf8KO]

ABSTRACT: To identify target genes of the histone demethylase Phf8 in oligodendroglial cells in a genome-wide and unbiased approach, RNA-sequencing studies were performed on wildtype CG4 cells and CRISPR/Cas9-edited Phf8 knockout cell lines. We compared three wildtype and one CRISPR/Cas9-treated clone without genome editing with four CRISPR/Cas9 Phf8-knockout clones. Among others we detected substantial changes in the expression of genes associated with cell cycle and replication of DNA.

ORGANISM(S): Rattus norvegicus

PROVIDER: GSE253877 | GEO | 2024/04/12

REPOSITORIES: GEO

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Genome-wide identification of target genes for the histone demethylase Phf8 in the oligodendroglial cell line CG4 [Phf8OE]

Project description:To identify target genes of the histone demethylase Phf8 in oligodendroglial cells in a genome-wide and unbiased approach, RNA-sequencing studies were performed on CG4 cells transduced with control or Phf8-overexpressing retroviruses. Transduced, GFP-expressing cells underwent fluorescence-activated cell-sorting, were expanded and then differentiated for three days. Two biological replicates were used for each condition. Gene expression analysis revealed an increase in oligodendrocyte progenitor proliferation and a general decrease in cell differentiation.

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Genome-wide identification of target genes for the transcription factors Nkx2.2 and Zfp488 in differentiating CG4 cells as a model for differentiating oligodendrocytes

Project description:In an attempt to validate the CG4 cell line as a model for oligodendrocytes, we determined the expression profiles in both cell types and the alterations during differentiation and found a striking overlap. Using CG4 cells as a model system for oligodendroglial cells, we generated several Nkx2.2 and Zfp488 overexpressing CG4 lines and compared their expression profile under differentiating conditions to regular CG4 cells to identify target genes of the two transcription factors in a genome-wide and unbiased approach. Several target genes were shared between Nkx2.2 and Zfp488, among them Gpr37.

2024-03-27 | GSE244592 | GEO

RNA-seq data of PHF8 wild-type and PHF8 knockout TRAMP mouse

Project description:RNA-seq analysis were applied to elucidate the transcriptional differences of PHF8 wild-type and PHF8 knockout TRAMP mouse. A total of 2,092 differentially expressed genes (Fold Change > 2, or Fold Change＜0.5; FDR < 0.05) with 623 down- and 1469 up-regulated genes were identifed in Phf8-KO TRAMP mice.

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Determination of Sox10-regulated genes in the oligodendroglial precursor cell line OLN93

Project description:To determine Sox10-regulated genes in the oligodendroglial precursor cell line OLN93, CRISPR/Cas9-mediated genome editing was performed to inactivate Sox10. For CRISPR/Cas9-mediated Sox10 inactivation, two guide sequences (5’-GCTGCGAGCCAGCCCAGGGCC-3’ for clone A and 5’-GTTGCCCAATTCGCCG GGCCC-3’ for clone B) were separately cloned into pX330. Oln93 cells were co-transfected with pEGFP-N1 and the pX330 plasmids that express the respective guide RNA and SpCas9. Transfected GFP-positive cells were enriched by fluorescence activated cell-sorting and seeded at single cell density in 96-well plates. Homogeneously GFP-expressing clones were analysed for loss of Sox10 protein by immunocytochemistry. Clones without Sox10 protein were obtained with both guide RNAs. Sox10 inactivation was confirmed by Sanger sequencing of the Sox10 gene. RNA was isolated from one clone A and one clone B as well as two independent RNA samples of wildtype OLN93 cells and submitted to RNA-seq.

2019-12-15 | GSE136659 | GEO

Analysis of transcriptional and epigentic changes induced by loss of Phf8 [RNA-seq]

Project description:Phf8 is a histone demethylase with links to human behavior. Here we have performed a transcriptional analysis of Phf8 knockout embryonic stem and neural projenitor cells in vitro along with an analysis of the prefrontal cortex and ventral striatum in vivo. Our results suggest that knockout of Phf8 is largely compensated for. However, we did observe a misregulation of serotonin receptors in the prefrontal cortex, which may have disease implications.

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Target of Ascl1 in CG4 cell

Project description:To identify the downstream targets of transcription factor Ascl1, transcriptional profiling of rat glial cell line, CG4 cell, transfected with either Ascl1 or mock control was performed. Comparison was performed between Ascl1 overexpressing CG4 cell and mock control transduced cell on 3D-Gene rat mRNA oligo 12k chip. Biological replication was not prepared. The results were confirmed by real time RT-PCR in several genes.

2010-12-01 | E-GEOD-22157 | biostudies-arrayexpress

RNA-seq data of PHF8 wild-type and PHF8 knockout TRAMP mouse

Project description:RNA-seq data of PHF8 wild-type and PHF8 knockout TRAMP mouse

| PRJNA662246 | ENA

Target of Ascl1 in CG4 cell

2010-12-01 | GSE22157 | GEO

APOC2 increase PHF8 binding on chromatin

Project description:We performed PHF8 ChIP-seq analysis of APOC2 overexpressed RKO cells and control cells to identify high-confident PHF8 regulated targets.

2021-05-07 | GSE114089 | GEO

The histone demethylase PHF8 is essential for cytoskeleton dynamics

Project description:PHF8 is a histone demethylase associated with X-linked mental retardation (XLMR). It has been described as a transcriptional coactivator involved in cell cycle progression, but its physiological role is still poorly understood. Here we show that PHF8 controls the expression of genes involved in cell adhesion and cytoskeleton organization such as RhoA, Rac1 and GSK3M-NM-2. A lack of PHF8 not only results in a cell cycle delay but also in a disorganized actin cytoskeleton and impaired cell adhesion. Our data demonstrate that PHF8 directly regulates the expression of these genes by demethylating H4K20me1 at promoters. Moreover, c-Myc transcription factor interacts with PHF8 and binds to the analyzed promoters, suggesting that c-Myc is involved on PHF8 recruitment to these promoters. Further analysis in the neuroblastoma cell line SH-SY5Y and in cortical neurons shows that depletion of PHF8 results in deficient neurite elongation. Overall, our results suggest that the mental retardation phenotype associated with loss of function of PHF8 could be due to abnormal neuronal connections as a result of alterations in cytoskeleton function. HeLa cells were transfected with either control or PHF8 siRNAs. Experiments were performed in biological triplicates. RNA were extracted and sujected to microarray analysis.

2012-07-31 | E-GEOD-38175 | biostudies-arrayexpress

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