Overexpression of Dmrt5 in in vitro differentiated neural progenitor cells regulates gene expression

To test the regulatory effects of Dmrt5 on gene expression, we designed tetracycline inducible lines of Dmrt5 transgenic mouse ESCs. Overexpression of Dmrt5 was induced upon addition of Doxycycline (Dox). To evaluate the effects of Dmrt5 on gene expression in different stages of in vitro differentiated NPC derived from mouse embryonic stem cells (ESC), we analyzed gene expression profiles at differentiation day 7 and day 9 with or without Dox. The data revealed that overexpression of Dmrt5 in in vitro differentiated neural progenitor cells (NPC) regulates gene expression. Addition of Dox to the medium of the control cell line rtTA did not significantly alter gene expression profile, demonstrating that the observed effects were through induction of Dmrt5, but not simply through Dox. A tetracycline-inducible rtTA-Dmrt5 ES cell line and the parental rtTA cell line were used for the experiment. These ES cells were differentiated using a monolayer differentiation (MD) protocol in N2B27 medium. At MD day 5, cells were treated with Dox or without. RNA was extracted from cells harvested at MD day 7 and day 9. Two replicates for each group were used for the experiment.