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

在之前的研究中，我们鉴定了NePro基因，这是一种在发育早期的小鼠新皮质中表达的基因，并表明NePro参与了Notch下游NPC的维持。在这项研究中，我们通过将缺失突变体导入鼻咽癌细胞来检测NePro的保守部分是否对其活性重要。与全长NePro相反，缺少NePro保守C末端部分的突变体并不抑制神经元分化，这表明该区域是NePro功能所必需的。此外，NLS被氨基酸取代打乱的NePro突变体没有表现出NePro活性，这表明NLS对NePro的功能是重要的。为了寻找在早期鼻咽癌中负责NePro表达的调控区域，我们构建了携带部分NePro基因组的报告载体。我们将它们电穿孔到胚胎小鼠的大脑中，并确定了位于NePro上游的一个调节区，该区域指导早期NPC中NePro的表达。

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.