Functional analysis of Nepro, a novel Notch downstream gene requiredfor the maintenance of neocortex neural progenitor cells

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In a previous study, we identified Nepro, a gene expressed in the developing mouse neocortex at early stages and showed that Nepro is involved in the maintenance of NPCs downstream of Notch. In this study, we examined whether the conserved parts of Nepro are important for its activity by transfecting deletion mutants into NPCs. In contrast to full-length Nepro, the mutant lacking conserved C-terminat part of Nepro did not inhibit neuronal differentiation, suggesting that this region is required for Nepro function. Furthermore, Nepro mutant in which the NLS is disrupted by amino acid substitutions did not exhibit Nepro activity, suggesting that the NLS is important for Nepro function. To search for regulatory regions responsible for Nepro expression in early NPCs, we constructed reporter vectors carrying parts of the Nepro genome. We electroporated them into the embryonic mouse brain and identified a regulatory region located upstream of Nepro, which directs Nepro expression in the early NPCs.