Project description:Differences in the amount of fetal hemoglobin (HbF) that persists into adulthood affect the severity of sickle cell disease and the beta-thalassemia syndromes. Genetic association studies have identified sequence variants in the gene BCL11A that influence HbF levels. Here we examine BCL11A as a potential regulator of HbF expression. The high HbF BCL11A genotype is associated with reduced BCL11A expression. Moreover, abundant expression of full-length forms of BCL11A is developmentally restricted to adult erythroid cells. Down-regulation of BCL11A expression in primary adult erythroid cells leads to robust HbF expression. Consistent with a direct role of BCL11A in globin gene regulation, we find that BCL11A occupies several discrete sites in the beta-globin gene cluster. BCL11A emerges as a therapeutic target for reactivation of HbF in beta-hemoglobin disorders. Expression clone label: FBB (4 different subclones, with 2 arrays each), Control label: MelBirA Experiment Overall Design: Microarray expression analysis from parental control mouse erythroleukemia (MEL) cells containing the BirA enzyme (MelBirA cells) and cells containing tagged versions (FLAG-Biotag) of BCL11A. Two control datasets and eight datasets from four subclones containing tagged BCL11A are included.
Project description:ChIP-seq analyses were performed in MEL cells expressing BirA alone or BirA and FLAG-Biotin tagged BCL11A (XL isoform). BCL11A chromatin occupancy in MEL cell line.
Project description:To promote the development and understanding of the in vitro erythroleukemia model, we analyzed the transcriptomes of mouse erythroleukemia (MEL) cells prior to and after erythroid-like differentiation induced by dimethyl sulfoxide (DMSO). A total of 348 protein-coding genes, including many known erythroid-enriched genes such as hemoglobin and heme synthesis genes, were upregulated upon erythroid-like induction in MEL cells
Project description:We compared the transcriptomes of ES cell derived erythroid progentor cells (ES-EP) and murine erythroleukemia (MEL) cells stably transfected with Gata-1 fused to ER. RNA was isolated from duplicate proliferating cultures of MEL and ES-EP using Affymetrix GeneChip Mouse Gene 1.0 ST.
Project description:We compared the transcriptomes of differentiating cultures of ES cell derived erythroid progentor cells (ES-EP) and murine erythroleukemia (MEL) cells stably transfected with GATA-1 fused to ER. RNA was isolated from duplicate differntiating cultures of MEL and ES-EP using Affymetrix GeneChip Mouse Gene 1.0 ST.
Project description:MEL clone 745 cells that are blocked at the pro-erythroblast stage serve as a model for terminal erythroid differentiation when treated with DMSO. A stable MEL cell line expressing (Dox)-dependent shRNA sequence targeting different regions of G9a mRNA was established and screened as previously described in (Demers, 2007 PMID17707229). Knock down was induced by adding 5 micrograms/ml final of Dox in the culture medium. For a total of six samples, 10 micro grams of RNA was extracted from each three biological replicates of mouse erythroleukemia (MEL) cells and their corresponding G9a knock downs. The RNA was extracted and purified using the RNeasy Mini Kit (Qiagen) including the on-column DNAse I digestion step. Experiment Overall Design: Mouse erythroleukemia cells: wild type vs G9 knock down. For each microarray hybridization, data analysis was performed using MAS and GCRMA.
Project description:Murine erythroleukemia (MEL) cells are differentiated by dimethyl sulfoxide (DMSO）, hexamethylene bisacetamide (HMBA） or trichostatin A (TSA） treatment. We selsected high differentiation-inducible (HD) and low differentiation-inducible (LD)-MEL cells by recloning of original MEL cells. We screened erythroid differentiation related-genes to compare transcriptome of HD and LD-MEL cells.
Project description:Murine erythroleukemia (MEL) cells are differentiated by dimethyl sulfoxide (DMSO), hexamethylene bisacetamide (HMBA), or trichostatin A (TSA) treatment. We analyzed expression profiles of high differentiation-inducible (HD) MEL cells during chemical induced differentiation.
