循环RNA（circRNA）是细胞过程的关键调节因子，在神经系统中丰富，对神经分化具有潜在的调节作用。然而，目前关于circRNA在脑发育中的功能的了解仍然有限。在这里，我们使用RNA测序技术展示了circRNA水平在人类胚胎干细胞分化为头端和尾端神经前体细胞（NPCs）的过程中显著增加，包括三个最丰富的circRNAs，即ciRS-7、circRMST和circFAT3。在神经分化的早期期间，circFAT3的敲除在大量RNA分析中导致轻微的转录变化。然而，在缺乏circFAT3的经过30和90天分化的人脑器官（cerebral organoids）的单细胞转录组学中，显示了前脑神经干细胞和成熟的皮层神经元的丢失。此外，cerebral organoids中的非前脑NPCs表达与神经分化和迁移相关的基因，包括FAT4、ERBB4、UNC5C和DCC的改变。在子宫内电穿孔小鼠前额叶皮层中，circFat3的体内消耗导致电穿孔细胞在新皮层内的位置发生变化。总体而言，这些发现表明circFAT3在神经发育中具有一致的作用，涉及前细胞类型的形成、神经元分化或迁移。©2023. 作者。

Circular RNAs (circRNAs) are key regulators of cellular processes, are abundant in the nervous system, and have putative regulatory roles during neural differentiation. However, the knowledge about circRNA functions in brain development is limited. Here, using RNA-sequencing, we show that circRNA levels increased substantially over the course of differentiation of human embryonic stem cells into rostral and caudal neural progenitor cells (NPCs), including three of the most abundant circRNAs, ciRS-7, circRMST, and circFAT3. Knockdown of circFAT3 during early neural differentiation resulted in minor transcriptional alterations in bulk RNA analysis. However, single-cell transcriptomics of 30 and 90 days differentiated cerebral organoids deficient in circFAT3 showed a loss of telencephalic radial glial cells and mature cortical neurons, respectively. Furthermore, non-telencephalic NPCs in cerebral organoids showed changes in the expression of genes involved in neural differentiation and migration, including FAT4, ERBB4, UNC5C, and DCC. In vivo depletion of circFat3 in mouse prefrontal cortex using in utero electroporation led to alterations in the positioning of the electroporated cells within the neocortex. Overall, these findings suggest a conserved role for circFAT3 in neural development involving the formation of anterior cell types, neuronal differentiation, or migration.© 2023. The Author(s).