Institute of Brain Science, National Yang-Ming University, Taipei, 112, Taiwan.
Department of Neurology, Neurological Institute, Taipei Veterans General Hospital, Taipei, 112, Taiwan.
Nat Commun. 2018 Jun 27;9(1):2498. doi: 10.1038/s41467-018-04880-8.
Mutations in genes involved in the production, migration, or differentiation of cortical neurons often lead to malformations of cortical development (MCDs). However, many genetic mutations involved in MCD pathogenesis remain unidentified. Here we developed a genetic screening paradigm based on transposon-mediated somatic mutagenesis by in utero electroporation and the inability of mutant neuronal precursors to migrate to the cortex and identified 33 candidate MCD genes. Consistent with the screen, several genes have already been implicated in neural development and disorders. Functional disruption of the candidate genes by RNAi or CRISPR/Cas9 causes altered neuronal distributions that resemble human cortical dysplasia. To verify potential clinical relevance of these candidate genes, we analyzed somatic mutations in brain tissue from patients with focal cortical dysplasia and found that mutations are enriched in these candidate genes. These results demonstrate that this approach is able to identify potential mouse genes involved in cortical development and MCD pathogenesis.
基因突变，尤其是与皮质神经元产生、迁移或分化相关的基因发生突变，往往会导致皮质发育异常（MCD）。然而，MCD 发病机制中涉及的许多基因突变仍未被识别。在这里，我们开发了一种基于转座子介导的体细胞核移植和胚胎电穿孔的体细胞诱变遗传筛选模型，并鉴定了 33 个候选 MCD 基因。与该筛选一致的是，其中一些基因已经涉及神经发育和疾病。候选基因的 RNAi 或 CRISPR/Cas9 功能破坏导致类似于人类皮质发育不良的神经元分布改变。为了验证这些候选基因的潜在临床意义，我们分析了局灶性皮质发育不良患者脑组织中的体细胞突变，发现这些突变在这些候选基因中富集。这些结果表明，这种方法能够识别潜在的与皮质发育和 MCD 发病机制相关的小鼠基因。
