Neuronal impact of patient-specific aberrant NRXN1α splicing [whole]

We performed targeted long-read and short-read RNA sequencing to identify and quantify NRXN1 isoforms in human post-mortem dlPFC and hiPSC-neurons derived from controls and NRXN1+/- individuals. We also performed whole transcriptome RNA seuqencing and 10x genomics single cell sequencing to determine transcriptional changes in NRXN1+/- hiPSC-neurons compared to controls. To investigate transcriptomic changes in NRXN1+/- neural cells, whole transcriptome short-read RNAseq was performed on hiPSC-NPCs and 6 week forebrain neurons (hiPSC-neurons) differentiated from multiple hiPSC clones derived from four control and four NRXN1+/- individuals totaling 40 unique samples. To examine the expression level of NRXN1 in the human brain, short-read RNAseq with three adult and one fetal human dlPFC sample. To examine specific hiPSC-derived neural cell types short-read RNAseq was performed on NGN2-induced neurons (3 weeks of differentiation) and ASCL1/DLX2-induced GABAergic neurons (5 weeks of differentiation) from control and NRXN1+/- donors. We also examined changes in cell type composition and neuronal maturation using 10X Genomics single-cell sequencing of two control hiPSC-neuron and three NRXN1+/- hiPSC-neuron samples.