Generation and analysis of disease model iPSCs. (A-B) Schematic diagram depicting the generation of patient-derived iPSCs by reprogramming (A) followed by the introduction of the DOX-inducible hADH5 expression vector into the hROSA26 locus by genome editing (B). (C-D) Positions of the primers (F4, F5, and R5) are indicated. Genomic PCR (C) and western blotting (WB) analysis (D) of the iPSC clones knocked in with the hADH5 expression vector. *Nonspecific band. (E) Generation of iPSCs with the indicated genotypes from 201B7 iPSCs by genome editing. The red dot in ADH5 exon 5 indicates a frameshift mutation created with genome editing (supplemental Figure 5). (F-G) Cell proliferation profile (F) and formaldehyde (HCHO) sensitivity (G) of the knockout iPSCs with the indicated genotypes. Mean ± standard deviation (SD) in triplicate cultures are shown. The experiment was repeated 3 times with similar results. (H) Cell cycle distribution of the patient-derived iPSCs, with or without DOX. Fixed cells were stained with propidium iodide and analyzed by a FACSCanto flow cytometer. Mean ± SD from 3 independent experiments are shown. (I-J) Cell proliferation profile (I) and formaldehyde (HCHO) sensitivity (J) of patient-derived iPSCs were analyzed with or without DOX-induced ADH5 expression. Mean ± SD in triplicate cultures are shown. The experiment was repeated 3 times with similar results.