Patient neural progenitors show a neurodevelopmental phenotype.

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Patient neural progenitors show a neurodevelopmental phenotype. Patient neural progenitors show a neurodevelopmental phenotype. (A) CRISPR/Cas9 targeting scheme for fluorescent PAX6-GFP reporter. (B) Southern blot analysis for targeted genomic locus showing 5′ external and internal probe for control and patient specific iPSC lines. (C) Representative immunostaining for GFP upon NPC differentiation. (D) FACS scheme separating three distinct cell populations. (E) Heat map of differentially expressed genes comparing GFP-positive, GFP-negative, and unsorted cell populations. (F) Scatter plot representing global gene expression levels. Blue dots represent up-regulated genes in GFP+ and GFP− fraction, respectively. Red dots highlight selected NPC marker genes in the GFP+ fraction. (G) GO term analysis of genes up-regulated in GFP+ fraction. (H) Quantitation of relative gene expression (rel. to CTRL NPCs) across patient-specific deletion. Selected NPC marker gene expression is shown as reference for PAX6-GFP sorted populations. (I) Differentially expressed genes comparing GFP+ cells form controls and patient line. (J) GO term analysis of genes down-regulated in patient NPCs (GFP+ cells). Reinhard Roessler et al. LSA 2018;1:e201800094 © 2018 Roessler et al.