Figure 1.

Establishment of CD-NS/PCs by genome editing and differentiation (A) schematic overview of genome editing. Some CD-NS/PCs were differentiated into neurons in vitro and subsequently utilized to assess the impact of 5-FC on neuronal cells. (B) Schematic depiction of the CRISPR/Cas9-mediated strategy for inserting the yCD-UPRT gene into the ACTB locus. Single guide RNA (sgRNA) target sequence and HR donor constructs are shown. (C) Cell viability of CD-NS/PCs at each 5-FC concentration. Micrographs of CD-NS/PCs cultured in the presence of 0, 7, 14, 35, or 50 μM of 5-FC for 7 days are also shown. Scale bar, 200 µm. (D) Relative mRNA expression of iPSCs, NS/PCs and neuron. (E) In vitro analysis of the effect of 5-FC on neurons. Nestin or beta III tubulin is labeled in green; Ki67 in red; and DAPI in blue. Ki67-positive cells remained without 5-FC, while they were absent in the presence of 5-FC, with beta III tubulin-positive neurons being preserved. Scale bar, 100 µm.