The ability of transcriptional regulators to drive lineage conversion of somatic cells offers great potential for the treatment of human disease. While current research in this field is focused on the generation of induced pluripotent stem cells or direct lineage transdifferentiation, less attention has been paid to the possibility of reprogramming cells to produce cytokines, growth factors and hormones. To explore the concept of switching on specific target genes in heterologous cells, we developed a model system to screen candidate factors for their ability to activate the archetypal megakaryocyte-specific chemokine platelet factor 4 (PF4) in fibroblasts. We found that co-expression of the transcriptional regulators GATA1 and FLI1 resulted in a significant increase in levels of PF4, which became magnified over time. We also determined that inclusion of a third factor, TAL1, further enhanced upregulation of PF4 expression. Our study therefore identified of TAL1 as an important component in the combination of transcriptional regulators that contribute to megakaryocyte programming, and demonstrated that such combinations can be used to produce potentially beneficial chemokines in readily available heterologous cell types.
Partial reprogramming of heterologous cells by defined factors to generate megakaryocyte lineage-restricted biomolecules.
Time
View SamplesKrppel-like factor 3 (KLF3) is a transcriptional repressor that has roles in adipogenesis, B-cell maturation and erythropoiesis (for review see Pearson et al., 2012).
Regions outside the DNA-binding domain are critical for proper in vivo specificity of an archetypal zinc finger transcription factor.
Specimen part
View SamplesTo investigate the roles of Klf3 in B lymphopoiesis, CD19+ B cells were sorted from the spleens of WT and Klf3 KO mice (Molecular and Cellular Biology (2008); 28:39673978).
Impaired B cell development in the absence of Krüppel-like factor 3.
Specimen part
View SamplesThe aim of this experiment was to investigate the regulation of gene expression by KLF3 and KLF8 in fetal erythroid cells by analyzing single and double mutant mouse models.
Generation of mice deficient in both KLF3/BKLF and KLF8 reveals a genetic interaction and a role for these factors in embryonic globin gene silencing.
Specimen part
View SamplesThe aim of this experiment was to investigate the role of KLF3 in regulating gene expression at different stages throughout the erythroid maturation process.
The CACCC-binding protein KLF3/BKLF represses a subset of KLF1/EKLF target genes and is required for proper erythroid maturation in vivo.
Specimen part
View SamplesThe aim of this experiment was to investigate the role of KLF1 in the fetal liver
The CACCC-binding protein KLF3/BKLF represses a subset of KLF1/EKLF target genes and is required for proper erythroid maturation in vivo.
Specimen part
View SamplesThe aim of this experiment was to investigate the dysregulation of gene expression in whole E12.5 embryos containing a gene trap (CH) or point mutation (H275R) within the Klf3 gene
ENU-induced mutation in the DNA-binding domain of KLF3 reveals important roles for KLF3 in cardiovascular development and function in mice.
Specimen part
View SamplesHuman diffuse intrinsic pontine gliomas (DIPG) are an aggressive form of pediatric brain tumors that arise in the pons in young children thus resulting in significant morbidity and very poor survival. Recent data suggest that mutations in the histone H3.3 variant are often found in these tumors, though the mechanism of their contribution to oncogenesis remains to be elucidated. Here we report that the combination of constitutive PDGFRA activation and p53 suppression as well as expression of the K27M mutant form of the histone H3.3 variant leads to neoplastic transformation of hPSC-derived neural precursors. Our study demonstrates that human ES cells represent an excellent platform for the modeling of human tumors in vitro and in vivo, which could potentially lead to the elucidation of the molecular mechanisms underlying neoplastic transformation and the identification of novel therapeutic targets.
Use of human embryonic stem cells to model pediatric gliomas with H3.3K27M histone mutation.
Specimen part
View SamplesSamples in this study probe the gene expression kinetics in human CCR6+ Th17 memory T cells activated under Th17 condition. Human CCR6+ Th17 memory T cells were purified from PBMC and gene expression was studied over a time course of 3 days after activation under Th17 condition. RNA from these samples was also profiled using RNA-Seq to compare different transcriptome profiling technologies.
Comparison of RNA-Seq and microarray in transcriptome profiling of activated T cells.
No sample metadata fields
View SamplesSeveral lipocalin genes from higher plants were shown to be responsive to both high and low temperature stresses and have been named as temperature-induced lipocalin (Til). In this study, a reverse genetic approach was taken to elucidate the role of Arabidopsis Til1 (At5g58070) in thermotolerance. We showed that Til1 proteins was constitutively expressed and increased significantly after heat shock treatment. A T-DNA knockout line of Til1, designated as til1-1, could not produce Til1 and showed severe defects in basal and acquired thermotolerance. Introducing a wild type copy of Til1 gene into til1-1 complemented the mutant phenotype. Over-expression of Til1 in the wild type plant did not enhance thermotolerance. Til1 is peripherally associated with plasma membrane, suggesting a regulatory or protective role of this protein in membrane function. Transcriptomic analysis showed that the heat shock response in til1-1 was not altered as compared to the wild type plants. The temperature threshold for heat shock protein induction was not affected by the level of Til1. Ion leakage analysis revealed no significant difference in membrane stability between the wild type and til1-1 seedlings. These results suggested that Til1 is not involved in regulating membrane fluidity or stability. Nevertheless, the level of malondialdehyde was significantly higher in til1-1 than in the wild type after severe heat treatment. The mutant plants were also more sensitive than the wild type to tert-butyl hydroperoxide, a reagent that induces lipid peroxidation. Taken together, our data indicate that Til1 is an essential component for thermotolerance probably by acting against lipid peroxidation induced by severe heat stress.
Temperature-induced lipocalin is required for basal and acquired thermotolerance in Arabidopsis.
No sample metadata fields
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