Single-cell RNA sequencing analysis of iPSC-derived motor neurons from an ALS patient carrying ATXN2 intermediate repeat expansions and a healthy control

Intermediate-length repeat expansions in ATAXIN-2 (ATXN2) are a strong genetic risk factor for amyotrophic lateral sclerosis (ALS). At the molecular level, ATXN2 intermediate expansions enhance TDP-43 toxicity and pathology. However, whether this triggers ALS pathogenesis at the cellular and functional level remains unknown. Here, we developed a human iPSC-derived model to investigate whether motor neurons derived from an ALS patient carrying ATXN2 intermediate repeat expansions (ALS-G) are transcriptomically distinct from a healthy control(OH2.6). For that, we generated a single cell RNA sequencing dataset consisting of a total of 384 cells. iPSC differentiation into MN was performed as described previously with minor modifications (Amoroso et al., 2013). Day 12 human iPSC-derived MN cultures were dissociated and sorted into single wells, where fluorescent-activated cell sorting (FACS) into 384-well plates was combined with automated single cell sequencing (SORT-seq pipeline). 384 cells were sorted into 4 libraries consisting of 96 cells each.