Over 1250 mutations in SCN1A, the Nav1.1 voltage-gated sodium channel gene, are associated with seizure disorders including GEFS+. To evaluate how a specific mutation, independent of genetic background, causes seizure activity we generated two pairs of isogenic human iPSC lines by CRISPR/Cas9 gene editing. One pair is a control line from an unaffected sibling, and the mutated control carrying the GEFS+ K1270T SCN1A mutation. The second pair is a GEFS+ patient line with the K1270T mutation, and the corrected patient line. By comparing the electrophysiological properties in inhibitory and excitatory iPSC-derived neurons from these pairs, we found the K1270T mutation causes cell type-specific alterations in sodium current density and evoked firing, resulting in hyperactive neural networks. We also identified differences associated with genetic background and interaction between the mutation and genetic background. Comparisons within and between dual pairs of isogenic iPSC-derived neuronal cultures provide a novel platform for evaluating cellular mechanisms underlying a disease phenotype and for developing patient-specific anti-seizure therapies.

中文翻译：

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两种同基因的人iPSC对的比较确定了由癫痫相关SCN1A突变直接引起的功能改变。

Nav1.1电压门控钠通道基因SCN1A中的1250多个突变与癫痫发作有关，包括GEFS +。为了评估独立于遗传背景的特定突变如何引起癫痫发作活动，我们通过CRISPR / Cas9基因编辑产生了两对同基因的人iPSC系。一对是来自未受影响兄弟姐妹的对照品系，是携带GEFS + K1270T SCN1A突变的突变对照。第二对是具有K1270T突变的GEFS +患者专线，以及校正后的患者专线。通过比较这些对中抑制性和兴奋性iPSC衍生神经元的电生理特性，我们发现K1270T突变会引起钠电流密度和诱发放电的细胞类型特异性变化，从而导致神经网络活跃。我们还确定了与遗传背景相关的差异，以及突变与遗传背景之间的相互作用。在双对同基因的iPSC衍生的神经元培养物中进行比较，从而为评估疾病表型所依据的细胞机制和开发针对患者的抗癫痫疗法提供了一个新颖的平台。