The evolutionarily conserved splicing regulator neuro-oncological ventral antigen 1 (NOVA1) plays a key role in neural development and function. NOVA1 also includes a protein-coding difference between the modern human genome and Neanderthal and Denisovan genomes. To investigate the functional importance of an amino acid change in humans, we reintroduced the archaic allele into human induced pluripotent cells using genome editing and then followed their neural development through cortical organoids. This modification promoted slower development and higher surface complexity in cortical organoids with the archaic version of NOVA1. Moreover, levels of synaptic markers and synaptic protein coassociations correlated with altered electrophysiological properties in organoids expressing the archaic variant. Our results suggest that the human-specific substitution in NOVA1, which is exclusive to modern humans since divergence from Neanderthals, may have had functional consequences for our species’ evolution.

进化上保守的剪接调节神经肿瘤腹侧抗原 1 ( NOVA1 ) 在神经发育和功能中起着关键作用。NOVA1还包括现代人类基因组与尼安德特人和丹尼索瓦人基因组之间的蛋白质编码差异。为了研究氨基酸变化对人类功能的重要性，我们使用基因组编辑将古老的等位基因重新引入人类诱导的多能细胞，然后通过皮质类器官跟踪它们的神经发育。这种修饰促进了具有古老版本的NOVA1的皮质类器官的更慢的发育和更高的表面复杂性. 此外，突触标记物和突触蛋白协同作用的水平与表达古老变体的类器官中电生理特性的改变相关。我们的结果表明，自尼安德特人分化以来现代人类所独有的NOVA1中的人类特异性替代可能对我们物种的进化产生了功能性影响。