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Transcriptomic changes in Sin3A deficient CD4 T cells

ABSTRACT: Highly purified CD4+ T cells from Rosa26-CreERT2 +/- Sin3AF/F micewere then cultured in interleukin 7 (IL-7) in the absence or the presence of 4-hydroxytamoxifen (TAM) for 6 days to allow Cre expression in the absence of cell division or differentiation . Next, we compared the behavior of Sin3A sufficient and deficient T cells in Th17 skewing conditions. RNA was then isolated and sequenced for the analysis of differential expression of mRNA transcripts

ORGANISM(S): Mus musculus

PROVIDER: GSE196615 | GEO | 2023/03/20

REPOSITORIES: GEO

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Mtor knockout in PDAC cells

Project description:A pancreatic ductal adenocarcinoma cell line was isolated from a Pdx1-Flp, FSF-KRasG12D/+, Rosa26-CreERT2/tdEG, Mtor-lox/lox mouse. Cultured cells were treated with 4-Hydroxytamoxifen to induce CreERT2 mediated excision of floxed Exon 3 of Mtor.

2017-10-16 | GSE98860 | GEO

RNA-seq expression profiling of Suz12-deleted or wild-type organoids derived from basal or luminal mouse mammary epithelial cells.

Project description:Total RNA was extracted from organoids grown from single sorted luminal or basal populations from the mammary glands of Rosa26-creERT2 Suz12 flox female mice. Cultures were treated with 4-hydroxytamoxifen to induce deletion of Suz12.

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ATAC-Seq chromatin profiling of Suz12-deleted or wild-type organoids derived from basal or luminal mouse mammary epithelial cells

Project description:Fixed DNA was isolated from organoids grown from single sorted luminal or basal populations from the mammary glands of Rosa26-creERT2 Suz12 flox female mice. Cultures were treated with 4-hydroxytamoxifen to induce deletion of Suz12.

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SIN3A Acetylation Complex - Fam60a defines a variant Sin3a-Hdac complex in embryonic stem cells required for self-renewal

Project description:Sin3a is the central scaffold protein of the prototypical Hdac1/2 chromatin repressor complex, crucially required during early embryonic development for the growth of pluripotent cells of the inner cell mass. Here, we explore the endogenous composition of the Sin3a-Hdac complex in pluripotent embryonic stem (ES) and differentiated cells. To do this, we established an endogenous double immunoprecipitation strategy coupled with quantitative mass spectrometry (ENDIP-MS) allowing us to define the precise composition of the Sin3a complex in multiple cell types. We identify the Fam60a subunit as a key defining feature of a variant Sin3a complex present in ES cells, but not in differentiated cells. Fam60a co-occupies H3K4me3 positive promoters with Sin3a and is essential to maintain it on chromatin. Consistent with this, Fam60a depletion phenocopies the loss of Sin3a, leading to decreased proliferation, an extended G1-phase and the deregulation of genes associated with differentiation. Taken together, our data characterise Fam60a as an essential core subunit of a variant Sin3a complex in ES cells required to promote rapid proliferation and to prevent unscheduled differentiation.

2017-05-19 | PXD005464 | Pride

SIN3A-regulated LIF-responsive genes in MCF7 cells

Project description:Tyrosine phosphorylation is a hallmark for activation of Signal Transducer and Activator of Transcription (STAT) proteins, but their transcriptional activity also depends on other secondary modifications. Type I interferons (IFNs) can activate both the ISGF3 (STAT1:STAT2:IRF9) complex and STAT3, but with cell-specific, selective triggering of only the ISGF3 transcriptional program. Following a genome-wide RNAi screen, we identified the Sin3a complex as an important mediator of this STAT3 transcriptional repression. Sin3a directly interacts with the DNA-binding domain of STAT3 and alters its acetylation status. SIN3A silencing enhances recruitment of STAT3 and enhanceosome components to the SOCS3 promoter, resulting in histone hyperacetylation and enhanced transcription. Conversely, Sin3a is required for ISGF3-dependent gene transcription and for an efficient IFN-mediated antiviral protection against Influenza A and hepatitis C viruses. The Sin3a complex therefore acts as a context-dependent STAT1/3 transcriptional switch. MCF7 cells were transfected with 50nM Renilla luciferase (control) or SIN3A-specific siRNA. 72h later, cells were cultured for 4h in absence of FCS and left non-stimulated or stimulated with LIF (10ng/ml, 1h). Total RNA was extracted. For each of the 4 conditions, 3 biological replicates were included. Nevertheless, one sample (SIN3A_siRNA_LIF1h_rep3) was discarded. In total 11 samples were analyzed.

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Role of polycomb repressive complex 1 in the intestinal homeostasis [RNA-seq 2]

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2015-10-29 | GSE71948 | GEO

Sin3A mutant pancreatic islets

Project description:Sin3A mutant pancreatic islets

| PRJNA610630 | ENA

Transcriptomes of Neural Stem and Progenitor Cells from the subgranular zone (SGZ) of Wild Type (NTG) and 3xTg-AD mice at juvenile age

Project description:The goals of this study are to compare transcriptome profiling (RNA-seq) in wild-type (NTG) and 3xTg-AD mice, to determine the transcriptomal changes of neural stem and progenitor cells (NSPC) from the SGZ at early stages of Alzheimer's Disease in a mouse model. Methods: A tamoxifen (TAM)-inducible Nestin-CreERT2;ROSA26-EYFP reporter mouse line was cross-bred with NTG or 3xTG-AD mice to label NSPC upon TAM administration. YFP-positive NSPCs were sorted through FACS (Fluorescence-Activated Cell Sorting) 10 days after TAM from NTG; Nestin-CreERT2;ROSA26-EYFP and 3xTg-AD;Nestin-CreERT2;ROSA26-EYFP mice. RNA was extracted with RNeasy Micro Kit and deep-sequenced on the NovaSeq SP flowcell SR200 platform. The sequence reads of Fastq files that passed quality filters were aligned to the mouse genome (build mm10) using hisat2. Samtools was used to index the BAM files. DESeq2 was used to establish differential gene expression between NTG and 3xTg-AD samples. Results: We mapped about 48 million sequence reads per sample to the mouse genome (build mm10) and identified 1391 genes that were differentially expressed in the 3xTg-AD, with 720 down- and 671 up-regulated. Genes were considered significantly different between two genotypes if they meet the following criteria: 1) adjusted p-value < 0.05, and 2) log2 fold change > 0.5 or < -0.5. Conclusions: Gene ontology analysis of differentially expressed genes uncovered several pathways that may contribute to early neural stem cell decline in the 3xTg mice. Our results revealed very early perturbations in the SGZ neurogenic niche in the 3xTg mouse model of AD, manifested by intrinsic molecular changes in neural stem and progenitor cells (NSPC) at juvenile age, resulting in dysregulated neural stem cell homeostasis.

2023-02-15 | GSE218323 | GEO

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