a, Schematic overview of the strategy to precisely modify the PLN sequence using CRISPR/Cas9 and single-stranded donor oligonucleotides (ssODN) complementary to the guide RNA (gRNA).
b, Sanger sequencing analysis showing the correction and introduction of the R14del variant sequence in hiPSCs generated from DCM and healthy individual hiPSCs, respectively.
c, Schematic representation of hiPSC lines used in the study.
d-e, Assessment of force generation of hiPSC-CMs carrying the PLN R14del mutation and their corresponding isogenic controls in 3D-EHTs, (2 batches, n = 5-12 EHTs each). f-g, 2D monolayer contractility measurements of hiPSC-CMs carrying the PLN R14del mutation and their corresponding isogenic controls (12 batches, n = 5-10 wells each). Colors represent experimental batches.
Abbreviations: HDR, homology-directed repair; EHTs, engineered heart tissues. Data were presented as mean ± SEM. * p < 0.05, ** p < 0.005, *** p < 0.0005.