The C9ORF72 Repeat Epansion Impairs Neurodevelopment

RNA sequencing analysis of human iPSC-derived neural stem cells generated from Control, C9ORF72 FTD/ALS, an isogenic control, and a C9ORF72 knockout line. The goal of this study is to determine the effects of the C9ORF72 repeat expansion on neural stem cell proliferation and differentiation. Neural stem cells were generated by exposing iPSCs to dual smad inhibition using a 10 day protocol. The neural stem cell population was enriched at day 10 using magnetic activated cell sorting (MACS) purification of polysialic acid-neural cell adhesion molecule (PSA-NCAM)+. The enriched population was lysed in RLT buffer and sent to Amaryllis Nucleics for 3'DGE RNA sequencing.

本研究针对源自四类样本的人类诱导多能干细胞（iPSC）衍生神经干细胞开展RNA测序分析，四类样本分别为对照组、C9ORF72相关额颞叶痴呆/肌萎缩侧索硬化（C9ORF72 FTD/ALS）组、同基因对照组以及C9ORF72基因敲除细胞系。本研究旨在明确C9ORF72基因重复扩增对神经干细胞增殖与分化的影响。实验采用为期10天的双Smad信号通路抑制方案处理诱导多能干细胞以诱导生成神经干细胞；于诱导第10天，通过磁激活细胞分选（MACS）纯化多唾液酸-神经细胞黏附分子（PSA-NCAM）阳性细胞，以此富集神经干细胞群体。将富集得到的细胞群体置于RLT缓冲液中裂解后，送至Amaryllis Nucleics进行3'端数字基因表达（3'DGE）RNA测序。