An in vivo model for analysis of developmental erythropoiesis and globin gene regulation.
Details
Publication Year 2014-01-17,Volume 28,Issue #5,Page 2306-7
Journal Title
FASEB Journal
Publication Type
JOURNAL ARTICLE
Abstract
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.-McColl, B., Kao, B. R., Lourthai, P. Chan, K., Wardan, H., Roosjen, M., Delagneau, O. Gearing, L. J., Blewitt, M. E, Svasti, S., Fucharoen, S., Vadolas, J. An in vivo model for analysis of developmental erythropoiesis and globin gene regulation.
Publisher
FASEB
Keywords
BAC transgenics, chromatin modification, epigenetics, hemoglobin switching
Research Division(s)
Molecular Medicine
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Copyright © 2014 by the Federation of American Societies for Experimental Biology


Creation Date: 2014-02-19 11:35:36
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