Pathogenic PPP2R5D variants alters the transcriptome of patient iPSC-derived glutamatergic neurons

The pathogenic PPP2R5D E198K and E420K variant alters the transcriptome of patient iPSC-derived cortical neurons Patient-iPSC derived iNGN2 glutamatergic neurons and isogenic controls were collected from three independent differentiations for each line (corE198K, E198K, corE420K, E420K) at DIV14, and total mRNA was extracted. cDNA libraries were made for each individual neuronal sample using Illumina TruSeq Stranded mRNA Library Prep Kit and 20 million pair-end reads were sequenced on an Illumina NovoSeq 6000.

致病性PPP2R5D E198K与E420K变异体可改变患者诱导多能干细胞（induced pluripotent stem cell, iPSC）来源皮层神经元的转录组。本研究针对各细胞系（corE198K、E198K、corE420K、E420K），收集其患者iPSC诱导的iNGN2谷氨酸能神经元及同基因对照样本；每组样本均通过三次独立分化获得，于体外培养第14天（days in vitro 14, DIV14）收集后提取总mRNA。随后采用Illumina TruSeq Stranded mRNA Library Prep Kit为每份神经元样本构建cDNA文库，并在Illumina NovaSeq 6000测序平台上完成2000万条双端读段的测序。