Gene expression profile of control and NGB knockdown human cerebral organoids at day 65.

We use single cell RNA sequencing (scRNA-seq) to analyze the impact of NGB on early neural development based on cerebral organoids model. Overall design: Three-dimensional cultured human cerebral organoids, derived from human induced pluripotent stem cells (hiPSCs), possess the remarkable capability to reconstitute in vivo three-dimensional cell arrangements. Moreover, they can faithfully recapitulate functional neural activity and gene expression profiles akin to those observed in the human brain. In this study, we employed lentiviral vectors to transduce short hairpin RNA (shRNA) targeting the NGB sequence, thereby generating an hiPSC line with NGB knockdown. Subsequently, this hiPSC line was utilized to generate cerebral organoids with NGB knockdown (shNGB) for future experimental analyses.