Project description:Sequencing of paediatric gliomas has identified two common substitution mutations (K27M and G34R) in genes encoding histone H3.3. We introduced a single-copy H3.3 G34R targeted mutation in mouse ES cells and observed gains in H3K36me3 and H3K9me3 across the genome. Altered chromatin profiles correlated with enrichment of KDM4 A/B/C, a histone lysine (K9/K36) demethylase. RNA-seq of H3.3 G34R mutant showed disrupted gene expression patterns which also correlated with KDM4 enrichment. Expression of a single copy of H3.3 G34R at endogenous levels was sufficient to genocopy KDM4 triple-KO cells as determined by ChIP-seq and RNA-seq.
Project description:Sequencing of paediatric gliomas has identified two common substitution mutations (K27M and G34R) in genes encoding histone H3.3. We introduced a single-copy H3.3 G34R targeted mutation in mouse ES cells and observed gains in H3K36me3 and H3K9me3 across the genome. Altered chromatin profiles correlated with enrichment of KDM4 A/B/C, a histone lysine (K9/K36) demethylase. RNA-seq of H3.3 G34R mutant showed disrupted gene expression patterns which also correlated with KDM4 enrichment. Expression of a single copy of H3.3 G34R at endogenous levels was sufficient to genocopy KDM4 triple-KO cells as determined by ChIP-seq and RNA-seq.
Project description:RNA-sequencing of H3.3-G34R and H3.3-WT HGG cells was performed to uncover transcriptomic differences related to the presence of H3.3-G34R mutation in a pediatric high grade glioma model, using a Sleeping beauty derived genetically engenieered mouse model.
Project description:ChIP-sequencing of H3.3-G34R and H3.3-WT HGG cells was performed for several histone marks to uncover differences on histone mark deposition related to the presence of H3.3-G34R mutation, using a Sleeping beauty derived genetically engenieered mouse model.
Project description:RNA-sequencing of H3.3-G34R and H3.3-WT HGG cells was performed to uncover transcriptomic differences related to the presence of H3.3-G34R mutation, using human cells obtained from a patient harboring pHGG and stably tranfected with 3X-FLAG-tagged wild type H3.3 or a 3X-FLAG-tagged H3.3 harboring the G34R mutation.
Project description:Regions of H3.3 binding in WT and ATRX KO mouse ES cells were identified by ChIP seq Chip-seq experiements were performed in WT and ATRX KO E14 mouse ES cells
Project description:The serine 31 (S31) residue of histone H3.3 is phosphorylated during mitosis and this modification is enriched at repetitive DNA. ChIP-seq of H3.3 S31Ph was carried out in a mitotic population of synchronised mouse ES cells. We find that H3.3 S31Ph is enriched at telomeres and IAP repeats in mitotic mouse ES cells.
Project description:This experiment was designed to study the effect of the histone H3.3-G34R mutation on the transcriptome of tumor cells and immune-infiltrating cells, in a high grade glioma mouse model.