Project description:Disruptor of telomeric silencing 1-like (DOT1L) is the specific and sole methyltransferase for H3K79. However, the role of Dot1l in T cells is not well determined. Here, used CD4-Cre/Dot1lflox/flox mice to specifically delete DOT1L in T cells. The genes characteristics were determined by RNA array and compared to wild type cells. We used microarrays to detail the global gene expression in Dot1l knockout and wild type CD8+ T cells.
Project description:We studied the role of the histone methyltransferase DOT1L in T cell development and differentiation. Here we generated RNA-seq data from sorted CD8 subpopulations from WT (Lcre+/-) and Dot1L KO (Lcre+/-;Dot1Lfl/fl) mice. This data was to define tissue specific expression signatures and it was compared with H3K79me2 ChIP-seq data from the same subsets.
Project description:DOT1L as methyltransferase of H3K79 is implicated in brian development. Here, we further defined DOT1L function in gene expression during cerebellar development using Microarrays. For that we generated Dot1l knockout mice using a Atoh-Cre driver line resulting in a Dot1l knockout within the cerebellum. The RNA of cerebellar tissue of the Dot1l knockout animals was thereby compared to controls. Additionally we compared the RNA levels of cultured CGNP and CGN samples treated with a DOT1L inhibitor versus DMSO treated cells. The data sets reveals potential new gene expression targets of DOT1L in vivo and in vitro, which ensure a correct development of the cerebellum.
Project description:to compare the transcriptomes of naïve CD4+ and CD8+ T-cells from Gfi1-knockout cells with wildtype cells, spleenocytes from wildtype and Gfi-knockout C57Bl/6 mice were isolated, stained with CD44, CD4 and CD8 antibodies and separated by using a FACS Diva (Becton Dickinson). Total RNAs isolated from 2 mice each (approximately 2xE06 cells) were pooled and subjected to microarray analysis on Affymetrix MOE430A_2.0 arrays Experiment Overall Design: 4 samples, 1 replicate per group
Project description:We studied the role of the histone methyltransferase DOT1L in T cell development and differentiation. We generated RNA-seq data from sorted CD8 effector T cells from WT (Lck-cre+/-) and Dot1L KO (Lck-cre+/-;Dot1Lfl/fl) mice infected with Listeria monocytogenes. This data was compared with H3K79me2 ChIP-seq data from the same T cell subset.
Project description:To investigate the function miR-144/451 on the function of CD8+ cells, we sorted the CD8a+ cells from wild type and miR-144/451 knockout mice, and then performed gene expression profiling analysis using data obtained from RNA-seq.
Project description:Many repetitive DNA elements are packaged in heterochromatin, but depend on occasional transcription to maintain long-term silencing. The factors that promote transcription of repeat elements in heterochromatin are largely unknown. Here, we show that DOT1L, a histone methyltransferase that modifies lysine 79 of histone H3 (H3K79), is required for transcription of major satellite repeats to maintain pericentromeric heterochromatin (PCH), and that this function is essential for preimplantation development. DOT1L is a transcriptional activator at single-copy genes but does not have a known role in repeat element transcription. We show that H3K79me3 is specifically enriched at repetitive elements, that loss of DOT1L compromises pericentromeric major satellite transcription, and that this function depends on interaction between DOT1L and the chromatin remodeler SMARCA5. DOT1L inhibition causes chromosome breaks and cell cycle defects, and leads to embryonic lethality. Together, our findings uncover a vital new role for DOT1L in transcriptional activation of heterochromatic repeats.