Department of Neurology, State Key Laboratory of Medical Neurobiology MOE Frontiers Center for Brain Science, Institutes of Brain Science, Zhongshan Hospital, Fudan University, Shanghai, 200032, China.
Department of Toxicology, School of Public Health, Fudan University, Shanghai, 200032, China.
J Neurosci. 2020 Sep 9;40(37):7169-7186. doi: 10.1523/JNEUROSCI.1044-20.2020. Epub 2020 Aug 12.
Conditional gene inactivation and restoration are powerful tools for studying gene functions in the nervous system and for modeling neuropsychiatric diseases. The combination of the two is necessary to interrogate specific cell types within defined developmental stages. However, very few methods and animal models have been developed for such purpose. Here we present a versatile method for conditional gene inactivation and restoration through reversibly inverting a critical part of its endogenous genomic sequence by Cre- and Flp-mediated recombinations. Using this method, we generated a mouse model to manipulate , an X-linked dosage-sensitive gene whose mutations cause Rett syndrome. Combined with multiple Cre- and Flp-expressing drivers and viral tools, we achieved efficient and reliable inactivation and restoration in the germline and several neuronal cell types, and demonstrated phenotypic reversal and prevention on cellular and behavioral levels in male mice. This study not only provides valuable tools and critical insights for and Rett syndrome, but also offers a generally applicable strategy to decipher other neurologic disorders. Studying neurodevelopment and modeling neurologic disorders rely on genetic tools, such as conditional gene regulation. We developed a new method to combine conditional gene inactivation and restoration on a single allele without disturbing endogenous expression pattern or dosage. We applied it to manipulate , a gene residing on X chromosome whose malfunction leads to neurologic disease, including Rett syndrome. Our results demonstrated the efficiency, specificity, and versatility of this new method, provided valuable tools and critical insights for function and Rett syndrome research, and offered a generally applicable strategy to investigate other genes and genetic disorders.
条件性基因敲除和恢复是研究神经系统中基因功能和模拟神经精神疾病的有力工具。为了在特定的发育阶段研究特定的细胞类型，这两种方法的结合是必要的。然而，为此目的开发的方法和动物模型非常少。在这里，我们通过 Cre 和 Flp 介导的重组可逆地反转其内源基因组序列的关键部分，提出了一种用于条件性基因敲除和恢复的通用方法。使用这种方法，我们生成了一种用于操纵的小鼠模型，是一种 X 连锁剂量敏感基因，其突变导致雷特综合征。结合多种 Cre 和 Flp 表达驱动子和病毒工具，我们在生殖系和几种神经元细胞类型中实现了高效和可靠的敲除和恢复，并在雄性小鼠中在细胞和行为水平上证明了表型逆转和预防。这项研究不仅为和雷特综合征提供了有价值的工具和关键见解，而且还为破译其他神经障碍提供了一种普遍适用的策略。研究神经发育和模拟神经障碍依赖于遗传工具，例如条件性基因调控。我们开发了一种新方法，可以在不干扰内源性表达模式或剂量的情况下，在单个等位基因上组合条件性基因敲除和恢复。我们将其应用于操纵，这是一个位于 X 染色体上的基因，其功能障碍导致包括雷特综合征在内的神经疾病。我们的结果证明了这种新方法的效率、特异性和通用性，为功能和雷特综合征研究提供了有价值的工具和关键见解，并为研究其他基因和遗传疾病提供了一种普遍适用的策略。
