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. 2022 Jul 27;23(15):8313. doi: 10.3390/ijms23158313

Figure 2.

Figure 2

Genetic correction of BrS-hiPSCs by CRISPR/Cas9 genome editing. (A) Corrected hiPSCs were generated with a CRISPR guide RNA targeting the CACNB2 exon 4 and a single-stranded oligonucleotide (ssODN) for homology-directed repair. (B) Confirmation of genetic correction, assessed by Sanger sequencing of genomic DNA. (C) Patients’ and CRISPR-corrected hiPSC lines exhibited a typical human stem cell-like morphology. Scale bar: 100 μm. (D) Immunofluorescence staining for key pluripotency markers OCT4, NANOG and TRA1–60 in patients’ and corrected hiPSC lines. Nuclei were counter-stained with DAPI. Scale bar: 100 μm. (E) Purity of patient-specific and CRISPR-corrected hiPSC lines was evaluated by the flow cytometry analysis of pluripotency markers OCT4 and TRA1–60. Gray dots represent the negative controls.