Figure 6: Electrophysiological features of corrected-LQTS3/BrS iPSC-derived cardiomyocytes, as determined by patch-clamp analyses.
From: Embryonic type Na+ channel β-subunit, SCN3B masks the disease phenotype of Brugada syndrome
(A) Schematic illustrations of the SCN5A gene correction implemented in the LQTS3/BrS iPSCs using HDAdVs. HDAdV, helper-dependent adenoviral vector; HSV-tk, herpes simplex virus thymidine kinase gene cassette; PGK-Neo, neomycin-resistance gene cassette; white triangles, loxP sites; mCMV-β-gal,β galactosidase gene cassette; red line, exon 28 of SCN5A containing the E1784K mutation. (B) PCR analysis confirmed that recombination occurred at the SCN5A locus. Products of 311 bp, 314 bp, 277 bp or 375 bp (the footprint of the targeting vector included) and 13156 bp were obtained using primers a-d, c-e, b-f and g-d (red arrowheads in A), respectively. (C) Sequence analysis of the SCN5A genes in each iPSC line. (D) Representative current trace of baseline in corrected-LQTS3/BrS iPSC-derived cardiomyocytes. (E–G). The average and data plots of peak current, late current, and relative late current in LQTS3/BrS (n = 13), corrected-LQTS3/BrS iPSC-derived cardiomyocytes (n = 7) and control-iPSC-derived cardiomyocytes (n = 6). (H) Averaged normalized LQTS3/BrS (n = 13) and corrected-LQTS3/BrS (n = 7) window currents obtained with a 100 ms repolarizing negative voltage ramp from +20 and −100 mV (1.2 mV/ms), normalized to the peak current recorded in the same cell.
