Fig. 1

Generation of FEV reporter hPSC. A Schematic diagram for strategy of CRISPR-Cas9-mediated knockin of reporter cassette into human FEV locus. B Genotyping of the edited hPSC (H9) clones with the primer sets (5FR, 3FR, FR) to identify the edited homozygous clone. (The 5FR and 3FR primer sets spanned the nucleotides outside the homologous arm to the internal reporter/selection cassette, aiming to verify the integration of the insertion cassette into the FEV locus; the FR primer set, flanking approximately 1500 bp of the stop codon of WT FEV gene, was designed to distinguish between homozygous and heterozygous clones) C Representative Sanger sequencing chromatograms for the homozygous Clone#7. D Immunofluorescence staining of the pluripotency markers (OCT4, NANOG, SSEA4, Tra-1-60, Tra-1-81) for the FEV-EGFP reporter cell line (H9-Clone#7). E H&E staining for the FEV-EGFP reporter cell line (H9-Clone#7)-derived teratoma. F Karyotyping analysis for the FEV-EGFP reporter cell line (H9-Clone#7). Scale bar for D, E 100 μm