Gene expression data from livers of 3-month-old HNF4alpha knockout mice
ABSTRACT: HNF4alpha is a master regulator of hepatic differentiation. In this study, HNF4alpha was deleted in adult mice using a Cre-LoxP system where Cre recombinase was delivered using an AAV8 virus. Total RNA was isolated from the livers of HNF4alpha-floxed mice (mixed backgournd) treated with either Control Virus or Cre-carrying virus at 3 months of age.
Project description:In this study, we analyzed global liver gene expression in MICU1 knock-down (KD) mice. To generate liver-specific MICU1 KD mice, MICU1loxp/loxp male mice were treated with an AAV8-Cre under the control of a hepatocyte specific promoter (TBG). AAV8-TBG-Null treated littermates were used as controls. Liver samples were collected 3-5 weeks after injection. Knockdown was verified by protein and mRNA (94%, 98%, respectively). Mouse Gene 2.0 ST (Affymetrix, Santa Clara, CA) arrays were used to obtain global gene expression data. Gene expression was profiled in control and in liver-specific Micu1 knockdown mice liver samples; 4 biological replicates per condition.
Project description:Analysis of changes in gene expression following hepatocyte specific deletion of GATA4 and GATA6 in adult mice. Results showed that the subset of differentially expressed genes had liver specific ontologies. Total RNA isolated from hepatocytes of AAV8-Tbg-Cre injected GATA4,6 fl/fl mice was compared to total RNA isolated from AAV-Tbg-GFP injected GATA4,6 fl/fl mice.
Project description:Analysis of changes in gene expression following hepatocyte specific deletion of GATA4 in adult mice. Results showed that the subset of differentially expressed genes had liver specific ontologies. Total RNA isolated from hepatocytes of AAV8-Tbg-Cre injected GATA4 fl/fl mice was compared to total RNA isolated from AAV-Tbg-GFP injected GATA4 fl/fl mice.
Project description:Liver firbrosis model of hepatocyte-specific FOXA2 knockout mice. Adeno-associated virus AAV8-TBG-Control or AAV8-TBG-Cre was injected via the tail vein of FOXA2flox/flox (FOXA2f/f) mice 2 weeks prior to CCl4 administration. Hepatic fibrosis was induced by injection of CCl4 twice per week for 4 weeks. Overall design: Adeno-associated virus AAV8-TBG-Control or AAV8-TBG-Cre was injected via the tail vein of FOXA2flox/flox (FOXA2f/f) mice 2 weeks prior to CCl4 administration. Hepatic fibrosis was induced by injection of CCl4 twice per week for 4 weeks. The mice were sacrificed on the third day after the last injection of CCl4 (0.25ml/kg BW). Total RNA was extracted from primary hepatocytes.
Project description:MIF is a general mediator of inflammatory responses and is associated with several inflammatory diseases including atherosclerosis. Therefore MIF is currently investigated as a therapeutic target for atherosclerosis. Generation of AAV8 vectors expressing human MIF was initiated to overexpress human MIF in AAV8 target tissues (liver, heart, muscles) of C57/Bl6 to study biological function of MIF in vivo and to provide an animal model for testing of MIF antibodies. Aim of the gene expression profiling experiment: Livers of AAV8-CMV-MIF treated animals in order to identify established and potential novel target genes of MIF induced signalling
Project description:Liver-specific depletion of HDAC3 leads to liver steatosis (fatty liver), suggesting disregulation of lipid metabolism. This is correlated with changes in lipid metabolic gene expression. Livers depleted of HDAC3 were removed from 12 week old male HDAC3 fl/fl mice (loxP sites flanking exon 4 to 7 of the HDAC3 gene encoding the catalytic domain of HDAC3) one week after the injection of AAV2/8-Tbg-Cre virus. Livers from the HDAC3 fl/fl mice injected with AAV2/8-Tbg-GFP were used as normal controls. mRNA was extracted from 100mg mouse liver samples and hybridized to Affymetrix microarrays. For each group (HDAC3 depleted liver and normal liver), we have 5 samples from different mice.
Project description:Canonical Wnt signalling regulates the self-renewal of most if not all stem cell systems. In the blood system, the role of Wnt signalling has been subject of much debate, with positive and negative roles of Wnt signalling proposed for hematopoietic stem cells (HSC). As we have shown previously, this controversy can be largely explained by the effects of different dosages of Wnt signalling. What remained unclear however, was why high Wnt signals would lead to loss of reconstituting capacity. To better understand this phenomenon, we have taken advantage of a series of hypomorphic mutant Apc alleles resulting in a broad range of Wnt dosages in HSCs, purified those HSCs and performed whole genome gene expression analyses. Gene expression profiling and functional studies show that HSCs with APC mutations lead to high Wnt levels , enhanced differentiation and diminished proliferation, but have no effect on apoptosis, collectively leading to loss of stemness. Thus, we provide mechanistic insight into the role of APC mutations and Wnt signalling in HSC biology. As Wnt signals are explored in various in vivo and ex vivo expansion protocols for HSCs, our findings also have clinical ramifications. To investigate the effects of Wnt signals in hematopoietic cells, mice carrying floxed Apc or hypomorphic Apc mutants were crossed, LSK cells were isolated and treated with Cre IRES GFP gamma-retrovirus ex vivo, GFP+ cells were sorted and RNA expression was determined.
Project description:Transcription profiling of mouse CD4+ and CD8+ T cells extracted from GFP-Egr2 knockin (Egr2 Kin) and hCD2-Cre / Egr2loxP/loxP / Egr3-/- Egr2/3 DKO) mice 7 days after infection with vaccinia virus.
Project description:The transcriptome of the anterior handplates of mouse limb buds of Gli3 constitutive and Hoxa13-Cre Gli3 conditional mutant embryos was compared to the transcriptome of limb buds of wild type and Gli3 heterozygote embryos at embryonic day E11.75. All samples carried one copy of the Hoxa13-Cre allele.
Project description:HDAC3 is a repressor of lipogenic gene expression whereas nuclear SREBP1c is a potent activiator that is controlled by SCAP. Since these factors share many lipogenic gene targets, we sought to determine their molecular crosstalk by ChIP-sequencing. We also tested whether ectopic expression of nSREBP1c interferes with HDAC3 binding near lipognic genes. To faciliate these pulldowns, we ectoipcally expressed HA-tagged nSREBP1c in mouse livers in vivo using AAV8 viruses and the thyroxine-binding globulin promoter for hepatocyte-specifc expression. ChIP-sequencing identified over 7,000 HA-nSREBP1c binding sites, almost 50% of which were in the same vicinity as HDAC3 peaks. However, ectopic expression of nSREBP1c did not disrupt HDAC3 binding indicating that these factors indepedently bind near lipogenic genes and are regulated by distinct regulatory pathways. Gene expression changes in livers of HDAC3 liver specific knockout (LKO), SCAP LKO, and double LKO mice are provided. Overall design: C57Bl/6 mice were injected with adeno-associated virus serotype 8 (AAV8) viruses expressing either GFP (control), HA-nSREBP1c, or Cre. Anti-HDAC3 or anti-HA antibodies were used for ChIP. For inputs, mouse livers lacking HDAC3 were used for HDAC3 ChIPs, and mouse livers expressing GFP were used for HA ChIPs. For gene expression, mice injected with AAV8:GFP were used as controls and indicated floxed mice were injected with AAV8:Cre.