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Runx3 regulates effector CD8 T cell differentiation and enhancer landscape

ABSTRACT: Runx3 regulates effector CD8 T cell differentiation and enhancer landscape

PROVIDER: PRJNA322854 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR3586800.fastq.gz	Fastqsanger.gz
	SRR3586801.fastq.gz	Fastqsanger.gz
	SRR3586802.fastq.gz	Fastqsanger.gz
	SRR3586803.fastq.gz	Fastqsanger.gz
	SRR3586804.fastq.gz	Fastqsanger.gz

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Impact of Runx3 deficiency on the epigenome of early effector CD8 T cells

Project description:Define how ablating Runx3 affects histone marks and active enhancers in early effector CD8 T cells

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Runx3 function in IL-2-activated splenic CD8+ T cells

Project description:CD8+T cells are immune cells that recognize foreign antigens on infected and tumor cells, leading to cytokine-dependent expansion and activation of cytotoxicity towards the targets. To identify Runx3 regulated genes, CD8+ T cells were isolated from spleen of WT and Runx3-/- mice . Six samples (3 WT and 3 Runx3-/-) of CD8+ T cells were separately obtained from individual mice, TCR activated and cultured for 4 days with IL-2.

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Genome-wide maps of Runx3 bound regions in splenic CD8+ T cells.

Project description:ChIP-seq was conducted using freshly isolated (resting) splenic WT CD8+ T cells with anti-Runx3 antibody (Ab), anti-H3K4me1 Ab and non-immune serum (NIS) as control. Two biological Runx3 and two NIS IP repeats from splenic CD8+ T cells isolated by positive selection on anti-CD8 magnetic beads, collected from 18 WT mice.

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Genome-wide maps of Runx3 bound regions in splenic IL-2-activated CD8+ T cells

Project description:ChIP-seq was conducted on isolated splenic WT CD8+ T cells, TCR-activated and cultured with IL-2 using anti-Runx3 antibodies (Ab), anti-H3K4me1 Ab and non-immune serum (NIS) as control. Two biological Runx3 and two NIS IP repeats from TCR-activated and IL-2 cultured splenic CD8+ T cells isolated by positive selection on anti-CD8 magnetic beads.

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Genome wide map of Runx3 bound regions in murine splenic CD8 T cells

Project description:Runx3 is an important transcription factor for the proper development of CD8+T cells. The number and functionality of CD8+T cells is severely affected in the absence of Runx3. To gain insight into the transcriptional program managed by Runx3 in CD8+T cells we conducted ChIP-seq on 50 million cells and CUT&RUN on 150,000 cells, using anti Runx3 antibodies. Both assays revealed similar results.

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Canonical BAF complex activity shapes the enhancer landscape that licenses CD8+ T cell effector and memory fates

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The interactomes of RUNX1 and RUNX3 from Mus musculus T cells

Project description:Cellular binary fate decisions require the progeny to silence genes associated with the alternative fate. The major subsets of alpha:beta T cells have been extensively studied as a model system for fate decisions. While the transcription factor RUNX3 is required for the initiation of Cd4 silencing in CD8 T cell progenitors, it is not required to maintain the silencing of Cd4 and other helper T lineage genes. The other runt domain containing protein, RUNX1, silences Cd4 in an earlier T cell progenitor, but this silencing is reversed whereas the gene silencing after RUNX3 expression is not reverse. Therefore, we hypothesized that RUNX3 and not RUNX1 recruits other factors that maintains the silencing of helper T lineage genes in CD8 T cells. To this end, we performed a proteomics screen of RUNX1 and RUNX3 to determine candidate silencing factors.

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RNA-Seq of CD8+ T cells expressing different levels of Runx3 in during LCMV infection (in vivo)

Project description:T cell receptor (TCR) stimulation of naïve CD8+ T cells initiates reprogramming of cis-regulatory landscapes that specify effector and memory cytotoxic T lymphocyte (CTL) differentiation. We mapped regions of hyper-accessible chromatin in naïve cells during TCR stimulation and discovered that the transcription factor (TF) Runx3 controls de novo access to memory CTL-specific cistromes prior to the first cell division, and is essential for memory CTL differentiation. Runx3 specifically promotes accessibility of cis-acting regions highly enriched with IRF, bZIP and Prdm1-like family TF motifs, upregulates IRF4 and establishes feed-forward transcriptional circuits that induce fundamental CTL attributes in memory precursor cells. Runx3 drives uncoupling from the naïve cell state, but subsequently restrains terminal differentiation of nascent CTL by preventing high expression of the TF T-bet and slowing effector cell proliferation. Enforced Runx3 expression enhances memory CTL differentiation and increases their numbers during iterative infections. Thus, Runx3 functions in a pioneering role to initialize and then ensure memory CTL differentiate.

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