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Genome-wide profile of Mpp8- and Tasor-DNA binding in mouse embryonic stem cells (mESCs) [CUT&Tag]

ABSTRACT: Purpose: To determine the genome-wide binding sites of Mpp8 and Tasor in mESCs

ORGANISM(S): Mus musculus

PROVIDER: GSE305571 | GEO | 2025/11/13

REPOSITORIES: GEO

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Genome-wide profile of Mpp8- and Tasor-DNA binding in mouse embryonic stem cells (mESCs) [CUT&Tag]

Project description:Genome-wide profile of Mpp8- and Tasor-DNA binding in mouse embryonic stem cells (mESCs) [CUT&Tag]

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Epigenomic profiling of TASOR in Hela cells using CUT&RUN and CUT&Tag

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Next-Generation Sequencing Facilitates Quantitative Analysis of WT and Mpp8/Tasor double knockout Embryonic Stem Cells Transcriptomes

Project description:Purpose: The goals of this study are to compare Mpp8/Tasor-dko with wildtype embryonic stem cells transcriptome profiling (RNA-seq) to evaluate the target genes regulated by HUSH complex.

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TASOR expression in naive embryonic stem cells safeguards their developmental potential [CUT&TAG]

Project description:The seamless transition through stages of pluripotency relies on a delicate balance between transcription factor networks and epigenetic silencing mechanisms that ensure proper regulation of the developmental program, critical for normal development. Here, we uncover the pivotal role of the transgene activation suppressor (TASOR), a component of the human silencing hub (HUSH) complex, in sustaining cell viability during the transition from naive to primed pluripotency, despite its rapid downregulation during this transition. Loss of TASOR in naive cells triggers replication stress, disrupts H3K9me3 heterochromatin formation, and compromise the transcriptional and post-transcriptional silencing of LINE-1 (L1) transposable elements (TEs), with these effects become more pronounced in primed cells. Remarkably, the survival of Tasor-knockout cells during naive to primed transition can be restored through the inhibition of cysteine-aspartic acid protease (Caspase) or deletion of mitochondrial antiviral signaling protein (MAVS). This suggests that unscheduled L1 expression activates an innate immune response, leading to programmed cell death, specifically in cells exiting naïve pluripotency. Additionally, we propose that HUSH-promoted H3K9me3 in naïve PSCs sets the stage for ensuing DNA methylation in primed cells, establishing long-term silencing during differentiation. Our findings shed insights on the crucial impact of epigenetic programs established in early developmental stages on subsequent phases, underscoring their significance in the developmental process.

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H3K9me3 ChIP-seq in HeLa cells lacking TASOR, MPP8, periphilin and SETDB1

Project description:We performed ChIP-seq to measure H3K9me3 levels in wild-type HeLa cells and HeLa cells lacking TASOR, MPP8, periphilin and SETDB1 generated through CRISPR/Cas9-mediated gene disruption.

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TASOR epigenetic repressor cooperates with a CNOT1 RNA degradation pathway to repress HIV

Project description:The Human Silencing Hub (HUSH) complex constituted of TASOR, MPP8 and Periphilin is involved in the spreading of H3K9me3 repressive marks across genes and transgenes such as ZNF encoding genes, ribosomal DNAs, retroelements, including LINE-1 elements, or the integrated HIV provirus. The deposit of these repressive marks leads to heterochromatin formation and inhibits gene expression. The precise mechanisms of HUSH-mediated silencing are still poorly understood, but HUSH presents structural homology with the RNA-induced transcriptional silencing (RITS) complex found in fission yeast. During transcription elongation by RNA polymerase II, RITS recruits a TRAMP-like RNA degradation complex composed of CNOT1 partners, MTR4 and the exosome, to ultimately repress gene expression via H3K9me3 deposit. Here, we show that TASOR depletion or HIV-2 Vpx expression, which induces TASOR degradation, increases the accumulation of transcripts derived from the HIV-1 LTR promoter at a post-transcriptional level. Furthermore, using a yeast 2-hybrid screen, we identified new TASOR partners involved in RNA metabolism including the RNA deadenylase CCR4-NOT complex scaffold CNOT1. TASOR and CNOT1 interact in vivo and synergistically repress HIV expression from its LTR. Similar to the RITS complex, we show that TASOR interacts and cooperates with MTR4 and the exosome. We also highlight an interaction between TASOR and RNA Polymerase II, predominantly under its elongating state, and between TASOR and some HUSH-targeted nascent transcripts. Finally, we show that TASOR overexpression facilitates the association of the aforementioned RNA degradation proteins with RNA polymerase II and is detected at transcriptional centers. Altogether, we propose that HUSH operates at the transcriptional and post-transcriptional levels to repress HIV proviral gene expression.

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Project description:Cut&Tag using antibodies against H3K27me3, H3K4me1, H3K4me3, HeK27ac, IgG Cells: mouses embryonic stem cells (mESCs), RGd2 Culture conditions: 2i, 30hr N2B27/DMSO/siNegative, 30hr N2B27MEK(i)/siNanog

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