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KCNQOT1 ChIRP seq and perturbation epigentic profiles [MeDIP]

ABSTRACT: KCNQOT1 ChIRP seq and perturbation epigentic profiles [MeDIP]

PROVIDER: PRJNA858013 | ENA |

REPOSITORIES: ENA

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KCNQOT1 ChIRP seq and perturbation epigentic profiles [MeDIP]

Project description:To study the molecular function of lncRNA KCNQOT1, we performed ChIRP-seq HEK293T cells, and we knocked it down in NIH3T3 and HEK293T cells, and created knockout of the repeat-rich and non-repeat regions of KCNQOT1 separately in HEK293T cells, then performed H3K9me3 ChIP-seq, DNA methylation MeDIP-seq and RNA-seq in the perturbed and control wild type cells.

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Project description:Transposon (de)repression and heterochromatin reorganization are dynamically regulated during cell fate determination and are hallmarks of cellular senescence. However, whether they are sequence specifically regulated remains unknown. Here, we uncover the KCNQ1OT1 lncRNA, by sequence-specific Hoogsteen base-pairing with double-stranded genomic DNA via its repeat-rich region and binding to heterochromatin protein HP1⍺, guides, induces and maintains epigenetic silencing at specific repetitive DNA elements. Repressing KCNQ1OT1 or deleting its repeat-rich region reduces DNA methylation and H3K9me3 on KCNQ1OT1 targeted transponsons. Engineering a fusion KCNQ1OT1 with an ectopically targeting guiding triplex sequence induces de novo DNA methylation at the target site. Phenotypically, repressing KCNQ1OT1 induces senescence associated heterochromatin foci, transposon activation and retrotransposition, and cellular senescence, demonstrating an essential role of KCNQ1OT1 to safeguard against genome instability and senescence.

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A ChIRP-MS analysis of HOTAIR in HeLa

Project description:We employed ChIRP in combination with proteomic strategy to systematically discover HOTAIR-interacting proteins. Three independent biological replicates of ChIRP-MS experiment were performed, alongside negative controls.

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