Project description:Chip-seq study for the genome-wide identification of CBX7 binding sites in CBX7 overexpressing CD34+ cellChip-seq study for genome-wide mapping of H3K9me3 and H3K27me3 in CD34+ cord blood cells upon overexpression of Polycomb CBX7 and Polycomb CBX8
Project description:The goal of this study is to identify m6A targets in human CD34+ HSPCs with or without STM2457 treatment using DARTseq. Purified human CD34+ HSPC cells were transduced with APOBEC-YTH or empty vector control (MIG). 24hr post infection, CD34+ cells were treated with STM2457 at a concentration of 20µM for 2 days. GFP+ cells were sorted the next day. 3 replicates were performed.
Project description:The overall goal is to understand how does the DNA methylation Canyon interactions in CD34+ HSPCs can be impacted by the inhibition of EZH2
Project description:The overall goal is to understand how does the DNA methylation Canyon interactions in CD34+ HSPCs can be impacted by the inhibition of EZH2
Project description:By genome-wide ChIP-seq analyses we show that Cbx7 and Cbx8 share ~95% of their targets in bone marrow cells. Cbx7 and Cbx8 have only ~200 targets that are not overlapping between the two proteins. These distinct Cbx7 and Cbx8 targets show reciprocal expression patterns along hematopoietic differentiation.
Project description:By genome-wide ChIP-seq analyses we show that Cbx7 and Cbx8 share ~95% of their targets in bone marrow cells. Cbx7 and Cbx8 have only ~200 targets that are not overlapping between the two proteins. These distinct Cbx7 and Cbx8 targets show reciprocal expression patterns along hematopoietic differentiation. Overexpression of empty vector (negative control), flag-Cbx7 and flag-Cbx8 in 5-FU treated bone marrow cells (progenitor and stem cells), followed by 1 week culture and subsequent ChIP using flag-M2 agarose beads.
Project description:Polycomb proteins play an essential role in maintaining the repression of developmental genes in self-renewing embryonic stem cells. The exact mechanism allowing the derepression of polycomb target genes during cell differentiation remains unclear. Here, we show that several differentiation genes transiently recruit a Cbx8-containing Polycomb repressive complex (PRC) 1 during their early activation. Depletion of Cbx8 partially impairs the transcriptional activation of these genes. This correlates with a reduction in low but detectable levels of histone H3 lysine 27 acetylation. Prolonged gene activation results in eviction of PRC1 despite persisting H3K27me3. The composition of PRC1 is highly modular and changes when ES cells commit to differentiation. We further demonstrate that the exchange of Cbx7 for Cbx8 is required for the effective activation of differentiation genes. Taken together our results establish a function for a Cbx8-containing complex in facilitating the transition from a Polycomb-repressed chromatin state to an active state. As this affects several key regulatory differentiation genes this mechanism is likely to contribute to the robust execution of differentiation programs. Examination of cbx8 in ES E14 mouse cells in 2 condition before and after 72h stimulation with retinoic acid compared with IgG
Project description:Polycomb CBX7 was stably expressed in embryonal carcinoma cells in order to identify target genes. The study was specifically geared toward unveiling gene targets that are susceptible to DNA hypermethylation in adult cancer. A number of CBX7 target tumor suppressor genes were thereby identified and used in further studies to characterize the mechanism surrounding CBX7 mediated gene suppression. Keywords: CBX7 target gene identification A cDNA encoding the ORF of polycomb CBX7 under the control of EF1alpha promoter was transfected into the Tera-2 embryonal carcinoma cell line. Empty vector was used to transfect control cells. The cells were subject to stable selection using puromycin and a pooled population of CBX7 overexpressing cells was established. Gene expression analysis was performed by comparing the gene expression profile of CBX7 cells to empty vector control cells.
Project description:The overall goal is to understand how does the DNA methylation Canyon interactions in CD34+ HSPCs can be impacted by the inhibition of EZH2