Expresion profile of TGR-1 (Myc+/+) and HO15.19 (Myc-/-) infected with a retrovirus expressing Hhex or GFP (controls)
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ABSTRACT:
ORGANISM(S): Rattus norvegicus
SUBMITTER: Roque Bort
PROVIDER: E-GEOD-51467 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
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Project description:The aim of this experiment is to determine Hhex targets in the presence and absence of Myc.
2014-07-16 | GSE51467 | GEO
Project description:We have cloned and characterized a fusion gene NUP98/HHEX1 resulting from t(7;10) from a patient with acute myeloid leukemia (AML). As NUP98/HHEX acts as an aberrant transcriptional activator, putative targets were searched upon transient expression of the fusion in primary murine bone marrow cells. Experiment Overall Design: Murine bone marrow cells were transduced with a retrovirus (MSCV-IRES-GFP, MIG) expressing either NUP98/HHEX or NUP98/HOXA9 (or the empty vector), mRNA was isolated after 72h. Each experiment was performed in triplicates.
2008-06-11 | E-GEOD-10909 | biostudies-arrayexpress
Project description:The Hematopoietically-expressed homeobox (Hhex) transcription factor is overexpressed in human myeloid leukemias. Conditional knockout models of murine acute myeloid leukemia (AML) indicate that Hhex maintains leukemia stem cell self-renewal by enabling Polycomb-mediated epigenetic repression of the Cdkn2a tumor suppressor locus, encoding p16Ink4a and p19Arf. However, whether Hhex overexpression also affects hematopoietic differentiation is unknown. To study this, we retrovirally overexpressed Hhex in hematopoietic progenitors. This enabled serial replating of myeloid progenitors, leading to the rapid establishment of IL-3-dependent promyelocytic cell lines. Use of a Hhex-ERT2 fusion protein demonstrated that continuous nuclear Hhex is required for transformation, and structure function analysis demonstrated a requirement of the DNA binding and N-terminal repressive domains of Hhex for promyelocytic transformation. This included the N-terminal Pml interaction domain, although deletion of Pml failed to prevent Hhex-induced promyelocyte transformation, implying other critical partners. Furthermore, deletion of p16Ink4a or p19Arf did not promote promyelocyte transformation, indicating that repression of distinct Hhex target genes is required for this process. Indeed, transcriptome analysis showed that Hhex overexpression resulted in repression of several myeloid developmental genes. To test potential for Hhex overexpression to contribute to leukemic transformation, Hhex-transformed promyelocyte lines were rendered growth factor-independent using a constitutively active IL-3 receptor common b subunit (bcV449E). The resultant cell lines resulted in a rapid promyelocytic leukemia in vivo. Thus Hhex overexpression can contribute to myeloid leukemia via multiple mechanisms including differentiation blockade and enabling epigenetic repression of the Cdkn2a locus.
2018-01-30 | GSE109793 | GEO
Project description:Cardiac muscle differentiation in vivo is guided by sequential growth factor signals, including endoderm-derived diffusible factors, impinging on cardiogenic genes in the developing mesoderm. Previously, by RNA interference in AB2.2 mouse embryonic stem cells (mESCs), we identified the endodermal transcription factor Sox17 as essential for Mesp1 induction in primitive mesoderm and subsequent cardiac muscle differentiation. However, downstream effectors of Sox17 remained to be proven functionally. In this study, we used genome-wide profiling of Sox17-dependent genes in AB2.2 cells, RNA interference, chromatin immunoprecipitation, and luciferase reporter genes to dissect this pathway. Sox17 was required not only for Hhex (a second endodermal transcription factor) but also for Cer1, a growth factor inhibitor from endoderm that, like Hhex, controls mesoderm patterning in Xenopus toward a cardiac fate. Suppressing Hhex or Cer1 blocked cardiac myogenesis, although at a later stage than induction of Mesp1/2. Hhex was required but not sufficient for Cer1 expression. Over-expression of Sox17 induced endogenous Cer1 and sequence-specific transcription of a Cer1 reporter gene. Forced expression of Cer1 was sufficient to rescue cardiac differentiation in Hhex-deficient cells. Thus, Hhex and Cer1 are indispensable components of the Sox17 pathway for cardiopoiesis in mESCs, acting at a stage downstream from Mesp1/2. Keywords: Cardiac development, Embryonic stem cells, Endoderm, Myogenesis, RNA interference Genome-wide expression profiling of Sox17-dependent genes. Mouse embryonic stem cells expressing Sox17 or luciferase shRNA were differentiated for up to 10 days by the embryoid body method [PMID:8155574], then were analysed using Affymetrix microarrays. ESCs were transduced with lentiviral vectors coexpressing enhanced green fluorescent protein (eGFP) with shRNA against Sox17, or against firefly luciferase. Transduced cells were flow-sorted based on GFP fluorescence, grown as embryoid bodies, and transferred to tissue culture plates after 4.5 days [PMID:17360443]. Cells were harvested at days 0, 2, 4, 5, 6, 8 and 10 in two biological replicates, except where noted.
2015-07-30 | E-GEOD-71528 | biostudies-arrayexpress
Project description:Goals and objectives of this study: to identify genes of the Treg signature induced by consitutive expression of GARP or FOXP3 in antigen-specific Th cells with potential role for stabililization & maintenance of the regulatory program. Keywords: T-cell receptor stimulation, gene-regulation, comparative gene expression profiling, cell type comparison, human, regulatory T cells, FOXP3, GARP Human alloantigen-specific Treg cells (THU) and Th cells (CD4-39), established and described recently (Ocklenburg et al.. Lab Invest.2006; 86: 724-737), were sitmulated for 3 days with cognate antigen (EBV B cells) and IL2 as described (Ocklenburg et al.. Lab Invest.2006; 86: 724-737) and analyzed using human Affymetrix U133 2.0 in monoplicate. Th cells had been transduced with a retroviral vector containing human GARP (LRRC32) or FOXP3 and an IRES-driven GFP as marker or empty GFP control, sorted for GFP+, expanded as described recently (Ocklenburg et al.. Lab Invest.2006; 86: 724-737), and characterized functially, phenotypically, and genetically as described (WO/2007/113301). Abbreviations: Treg THU d3 = alloantigen-specific Treg cells; GFP d3 = GFP-transduced alloantigen-specific Th cells (CD4-39); FoxP3 d3 = FOXP-transduced alloantigen-specific Th cells (CD4-39); Garp d3 = GARP-transduced alloantigen-specific Th cells (CD4-39); all stimulated for 3 days (=d3) with EBV B cells and IL2.
2009-04-20 | E-GEOD-13234 | biostudies-arrayexpress
Project description:Expresion profile of TGR-1 (Myc+/+) and HO15.19 (Myc-/-) infected with a retrovirus expressing Hhex or GFP (controls)
| PRJNA223269 | ENA
Project description:Transcription profiling by array of wild type and p300 KIX/KIX; CBP +/KIX primary mouse embryonic fibroblasts (MEFs) transduced with either MSCV-c-Myb_IRES-GFP or MSCV-IRES-GFP retrovirus to determine the effect of mutating the KIX domain of CBP and p300 on c-Myb regulated gene expression. CBP KIX mutation (MGI:3578129) and p300 KIX mutation (MGI:3578128).
2014-02-10 | E-MTAB-1973 | biostudies-arrayexpress
Project description:MSCV-GFP-IRES (IRES), MSCV-GFP-Myc-NIC1 (NIC1) and MSCV-GFP-DNMAML (DNMAML) were retrovirally transduced in to THP-1 cell (pCL-Ampho was used as packaging plasmid). GFP positive cells were sorted for selective desired cell. Then IRES, NIC1 and DNMAML overexpressing THP-1 were pretreated with PMA (5ng/ml) for 48 h before stimulation with IL-4 (20 ng/ml) for 3 h.
2019-05-01 | GSE115914 | GEO
Project description:LSK cells transfected with Hhex retrovirus
| PRJNA432046 | ENA
Project description:Krüppel-like factor 1 (Klf1) is a transcription factor that promotes β-globin synthesis in erythroid progenitors, and its role in immunological homeostasis is unknown. The full-length mRNA of mouse Klf1(NM-010635) was cloned into the pMIG retroviral vector, which contains an IRES-GFP cassette. Splenocytes from C57BL/6 mice were cultured for 48 h in the presence of Concanavalin A (ConA) (10 μg/mL) and IL-2 (50 μg/mL), and gene transduction was conducted. CD4+ T cells that were transfected with retroviral vectors were divided into 3 fractions according to GFP fluorescence (strongly positive, weakly positive and negative) using a FACSVantage flow cytometer. Total RNA was then extracted. Sequence libraries were prepared according to standard methods using Smart-seq2 (Illumina), and they were analyzed with a next generation sequencer, a MiSeq system (Illumina).
2018-05-10 | E-MTAB-6778 | biostudies-arrayexpress