Figure 2.

Generation of Isogenic E99K hiPSC Cardiomyocytes

(A) A footprint-free PiggyBac targeting strategy of the ACTC1 gene was used to correct or induce E99K mutation into hiPSC lines.

(B–D) In (B), sequencing confirmed the (C)301G>A ACTC1 mutation had been corrected in E99K1 and E99K2 and introduced in NC. In (C), high-efficiency differentiation of hiPSC still yielded cardiomyocyte purities of >90% after gene editing (red is α-actinin staining; blue is DAPI). Counterstaining (green) with an antibody specific to the mutant E99K peptide confirmed protein expression was lost in diseased patients (C and D). In contrast, E99K reactivity was now detected in hiPSC cardiomyocytes from the healthy individual (C and D). n = 6. Scale bars, 50 μm.

Significance was determined by Student’s t test, ^∗∗∗p < 0.001.