naa a22 4500 528787047 20180924065502.0 cr unu---uuuuu 180924e20110728xx s 000 0 eng 10.3929/ethz-b-000038572 doi 10.1371/journal.pone.0022838 doi (ETHRESEARCH)oai:www.research-collection.ethz.ch:20.500.11850/38572 MicroRNA-96 Directly Inhibits gamma-Globin Expression in Human Erythropoiesis [Elektronische Daten] [Imane Azzouzi, Hansjoerg Moest, Jeannine Winkler, Jean-Claude Fauchère, André P. Gerber, Bernd Wollscheid, Markus Stoffel, Markus Schmugge, Oliver Speer] PLoS ONE Open access ethresearch Fetal hemoglobin, HbF (α2γ2), is the main hemoglobin synthesized up to birth, but it subsequently declines and adult hemoglobin, HbA (α2β2), becomes predominant. Several studies have indicated that expression of the HbF subunit γ-globin might be regulated post-transcriptionally. This could be confered by ∼22-nucleotide long microRNAs that associate with argonaute proteins to specifically target γ-globin mRNAs and inhibit protein expression. Indeed, applying immunopurifications, we found that γ-globin mRNA was associated with argonaute 2 isolated from reticulocytes that contain low levels of HbF (<1%), whereas association was significantly lower in reticulocytes with high levels of HbF (90%). Comparing microRNA expression in reticulocytes from cord blood and adult blood, we identified several miRNAs that were preferentially expressed in adults, among them miRNA-96. The overexpression of microRNA-96 in human ex vivo erythropoiesis decreased γ-globin expression by 50%, whereas the knock-down of endogenous microRNA-96 increased γ-globin expression by 20%. Moreover, luciferase reporter assays showed that microRNA-96 negatively regulates expression of γ-globin in HEK293 cells, which depends on a seedless but highly complementary target site located within the coding sequence of γ-globin. Based on these results we conclude that microRNA-96 directly suppresses γ-globin expression and thus contributes to HbF regulation. Creative Commons Attribution 3.0 Unported http://creativecommons.org/licenses/by/3.0 ethresearch Azzouzi Imane joint author Moest Hansjoerg joint author Winkler Jeannine joint author Fauchère Jean-Claude joint author Gerber André P. joint author Wollscheid Bernd joint author Stoffel Markus joint author Schmugge Markus joint author Speer Oliver joint author PLoS ONE Lawrence, KS, USA : Public Library of Science 6 (7), p. e22838 1932-6203 http://hdl.handle.net/20.500.11850/38572 text/html WWW-Backlink auf das Repository (Open access) 1 Journal Article ethresearch ETHRESEARCH 856 40 http://hdl.handle.net/20.500.11850/38572 text/html WWW-Backlink auf das Repository (Open access) ETHRESEARCH 700 1- Azzouzi Imane joint author ETHRESEARCH 700 1- Moest Hansjoerg joint author ETHRESEARCH 700 1- Winkler Jeannine joint author ETHRESEARCH 700 1- Fauchère Jean-Claude joint author ETHRESEARCH 700 1- Gerber André P. joint author ETHRESEARCH 700 1- Wollscheid Bernd joint author ETHRESEARCH 700 1- Stoffel Markus joint author ETHRESEARCH 700 1- Schmugge Markus joint author ETHRESEARCH 700 1- Speer Oliver joint author ETHRESEARCH 773 0- PLoS ONE Lawrence, KS, USA : Public Library of Science 6 (7), p. e22838 1932-6203 BK010053 XK010053 XK010000 ETHRESEARCH ETHRESEARCH ETHRESEARCH Journal Article Open access