An in vivo model for analysis of developmental erythropoiesis and globin gene regulation
| Autor: | Kasey Chan, Bradley McColl, Orane Delagneau, Linden J. Gearing, Suthat Fucharoen, Saovaros Svasti, Preeyachan Lourthai, Betty Kao, Mark Roosjen, Hady Wardan, Jim Vadolas, Marnie E. Blewitt |
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| Rok vydání: | 2014 |
| Předmět: |
Male
Time Factors Green Fluorescent Proteins Mutant Mice Transgenic beta-Globins Biology Biochemistry Epigenesis Genetic Green fluorescent protein Mice Erythroid Cells hemic and lymphatic diseases Genetics medicine Animals Erythropoiesis gamma-Globins Transgenes Epigenetics Yolk sac Promoter Regions Genetic DNA Modification Methylases Molecular Biology Gene Fetal Hemoglobin Gene knockdown Mesonephros Cell biology Mice Inbred C57BL Disease Models Animal medicine.anatomical_structure Gene Expression Regulation Mutation Female RNA Interference Biotechnology |
| Zdroj: | The FASEB Journal. 28:2306-2317 |
| ISSN: | 1530-6860 0892-6638 |
| Popis: | Expression of fetal γ-globin in adulthood ameliorates symptoms of β-hemoglobinopathies by compensating for the mutant β-globin. Reactivation of the silenced γ-globin gene is therefore of substantial clinical interest. To study the regulation of γ-globin expression, we created the GG mice, which carry an intact 183-kb human β-globin locus modified to express enhanced green fluorescent protein (eGFP) from the Gγ-globin promoter. GG embryos express eGFP first in the yolk sac blood islands and then in the aorta-gonad mesonephros and the fetal liver, the sites of normal embryonic hematopoiesis. eGFP expression in erythroid cells peaks at E9.5 and then is rapidly silenced (>95%) and maintained at low levels into adulthood, demonstrating appropriate developmental regulation of the human β-globin locus. In vitro knockdown of the epigenetic regulator DNA methyltransferase-1 in GG primary erythroid cells increases the proportion of eGFP(+) cells in culture from 41.9 to 74.1%. Furthermore, eGFP fluorescence is induced >3-fold after treatment of erythroid precursors with epigenetic drugs known to induce γ-globin expression, demonstrating the suitability of the Gγ-globin eGFP reporter for evaluation of γ-globin inducers. The GG mouse model is therefore a valuable model system for genetic and pharmacologic studies of the regulation of the β-globin locus and for discovery of novel therapies for the β-hemoglobinopathies. |
| Databáze: | OpenAIRE |
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