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Consequences of GATA1 expression in Gata1- G1ME Murine Megakaryocyte/Erythrocyte Progenitor Cell Line

ABSTRACT: G1ME cells are GATA1-deficient murine bipotential megakaryocyte/erythrocyte progenitor cells derived from Gata1-negative murine ES cells. In order to assess the impact of GATA1 on gene regulation and cell differentiation, an expression construct was used to transiently produce high levels of GATA1. Cells transduced with this construct or a vector control were harvested at 18 and 42 hours, and gene expression was analyzed using Affymetrix MOE430 version 2 arrays.

ORGANISM(S): Mus musculus

PROVIDER: GSE14980 | GEO | 2009/06/10

SECONDARY ACCESSION(S): PRJNA111885

REPOSITORIES: GEO

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Consequences of GATA1 expression in Gata1- G1ME Murine Megakaryocyte/Erythrocyte Progenitor Cell Line

Project description:G1ME cells are GATA1-deficient murine bipotential megakaryocyte/erythrocyte progenitor cells derived from Gata1-negative murine ES cells. In order to assess the impact of GATA1 on gene regulation and cell differentiation, an expression construct was used to transiently produce high levels of GATA1. Cells transduced with this construct or a vector control were harvested at 18 and 42 hours, and gene expression was analyzed using Affymetrix MOE430 version 2 arrays. Both vector control and GATA1-expressing cells were isolated by FACS for GFP and 18 and 42 hours. Biologic triplicates were performed for each construct at each timepoint.

2009-06-10 | E-GEOD-14980 | biostudies-arrayexpress

Genome-wide analysis of chromatin occupancy of GATA1 and its short isoform GATA1s (ChIP-Seq)

Project description:We report ChIP-Seq data for GATA1 and the leukemia-associated short isoform GATA1s in G1ME cells, a Gata1-null cell line with both erythroid and megakaryocytic differentiation potential. We introduced HA-tagged GATA1 or GATA1s into G1ME cells via retroviral transduction. The cells were crosslinked at 48h post-transduction, and an HA antibody was used for chromatin immunoprecipitation (ChIP). ChIP and input samples were sequenced on Illumina GAII or GAIIx high-throughput sequencers. The data reveal a deficiency of chromatin occupancy by GATA1s, especially at genes involved in erythrocyte differentiation and function.

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Genome-wide analysis of chromatin occupancy of GATA1 and its short isoform GATA1s (ChIP-Seq)

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2015-01-26 | GSE62879 | GEO

Differential gene regulation by the disease-associated short isoform of GATA1 (microarray)

Project description:The transcriptional activiy of GATA1s was compared to GATA1 through gene expression analysis in a cell line model with both erythroid and megakaryocyte differentiation. G1ME cells were derived from Gata1- mouse ES cells and have both megakaryocyte and erythrocyte differentiation potential upon reconstitution of GATA-1 expression (Stachura 2006). HA-tagged full length or short GATA-1 were expressed in G1ME cells grown in TPO via retroviral transductions. The cells were sorted for GFP positivity 68 hours post-transduction and then were allowed to recover in normal growth medium for 4h. Total RNA was then isolated using RNeasy kit from Qiagen 72 hours post-transduction.

2015-02-04 | E-GEOD-64494 | biostudies-arrayexpress

Transcriptome profiling of mouse Gata1- megakaryocyte-erythroid progenitors (G1MEs) expressing one of the two isoforms of GATA1: full-length (GATA1fl) or an amino-truncated form of GATA1 (GATA1s).

Project description:We transduced mouse Gata1- megakaryocyte-erythroid progenitors with MIGRI-GFP vector expressing GATA1fl or GATA1s cDNAs. GFP-positive cells expressing one of the two isoforms of GATA1 were isolated by FACS 42 hours following transduction and used for microarray transcriptome analysis. At this time point, there was no apparent difference in the cell surface phenotypes between GATA1fl and GATA1s-expressing cells. Transcriptome data for G1ME/GATA1fl at 42h were deposited previously under GSE14980 (GSM374049, GSM374050, GSM374051), whereas G1ME/GATA1s at 42h are deposited here. We performed differential expression analysis comparing mean expression levels of genes in G1ME cells at 42 hours after transduction with GATA1fl (3 replicates deposited previously under GSE14980: GSM374049, GSM374050, GSM374051) or GATA1s (3 replicates deposited here). All 6 samples were prepared the same way and analyzed on the same platform, Affymetrix Mouse Genome 420 2.0.

2015-01-26 | E-GEOD-62879 | biostudies-arrayexpress

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Project description:We explored the role of FOG-1 in GATA-1 transcriptional regulation of megakaryocyte differentiation through expression of wild-type GATA-1 and the FOG-binding mutant of GATA-1 (GATA-1^V205G) in G1ME cells. G1ME cells were derived from Gata1-null mouse ES cells and have both megakaryocyte and erythrocyte differentiation potential upon reconstitution of GATA-1 expression (Stachura 2006). HA-tagged wild-type or mutant GATA-1 were expressed in G1ME cells grown in TPO via retroviral transductions. The cells were sorted for GFP positivity 68 hours post-transduction and then were allowed to recover in normal growth medium for 4h. Total RNA was then isolated using RNeasy kit from Qiagen 72 hours post-transduction.

2012-03-05 | E-GEOD-35695 | biostudies-arrayexpress

Mus musculus

Project description:Consequences of GATA1 expression in Gata1- G1ME Murine Megakaryocyte/Erythrocyte Progenitor Cell Line

| PRJNA111885 | ENA

megamiR, microRNA expression profiles during megakaryocyte-erythrocyte progenitor (MEP) differentiation

Project description:This collection contains microRNA expression profiling data for samples purified from umbilical cord blood, belonging to one of the followiing populations: MEP, MEGA1, MEGA2, ERY1, ERY2, ERY3. MEP: megakaryocyte-erythrocyte precursors (defined by CD34+ CD38+ IL-3Ra- CD45RA- ); MEGA1: megakaryocyte population 1 (defined by CD34+ CD61+ CD41+ CD45- ); MEGA2: megakaryocyte population 2 (defined by CD34- CD61+ CD41+ CD45- ); ERY1: erythrocyte population 1 (defined by CD34+ CD71+ GlyA- ); ERY2: erythrocyte population 2 (defined by CD34- CD71+ GlyA- ); ERY3: erythrocyte population 3 (defined by CD34- CD71+ GlyA+ ). Keywords: microRNA, miRNA, MEP, megakaryocyte, erythrocyte, lineage specification

2008-06-10 | GSE10593 | GEO

Inducible Gata1 Suppression Expands Erythro-Megakaryocytic Progenitors from Embryonic Stem Cells

Project description:Embryonic stem (ES) and induced pluripotent stem (iPS) cells represent a potential source of megakaryocytes and platelets for transfusion therapies. However, most current ES/iPS cell differentiation protocols are limited by low yields of hematopoietic progeny. Mutations in the mouse and human genes encoding transcription factor GATA1 cause accumulation of proliferating, developmentally arrested megakaryocytes. To exploit this clinical observation, we engineered wildtype (WT) murine ES cells to express doxycycline (dox)-regulated Gata1 short hairpin (sh) RNAs. In vitro differentiation with dox and thrombopoietin (Tpo) resulted in approximately 1013-fold expansion of immature hematopoietic progenitors. Upon dox withdrawal with multilineage cytokines, GATA1 expression was restored and the cells differentiated into erythroblasts and megakaryocytes. With Tpo alone, dox-deprived progenitors formed mainly mature megakaryocytes that generated functional platelets in vivo. Our findings provide a novel, readily reproducible strategy to expand ES-cell derived megakaryocyte-erythroid progenitors and direct their differentiation into megakaryocytes producing functional platelets in clinically relevant numbers. 3 classes of samples were compared 1) fetal liver derived megkaryocytes 2) G1ME (Gata1â megakaryocyte-erythroid) 3) G1ME2 (engineered wildtype (WT) murine ES cells to express doxycycline (dox)-regulated Gata1 short hairpin (sh) RNAs)

2015-12-09 | E-GEOD-66479 | biostudies-arrayexpress

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