249 Unfolded Protein Response in PLN R14del Cardiomyopathy 9 Figure 1. Phospholamban (PLN) loss of arginine at position 14 (R14del) human induced pluripotent stem cell (hiPSC) disease modeling and functional assessment of contractility. A, Schematic overview of the strategy to precisely modify the PLN sequence using CRISPR/Cas9 and single-stranded donor oligonucleotides complementary to the guide RNA (gRNA). B, Sanger sequencing analysis showing the correction and introduction of the R14del variant sequence in hiPSCs generated from dilated cardiomyopathy and healthy individual hiPSCs, respectively. C, Schematic representation of hiPSC lines used in the study. D and E, Assessment of force generation of hiPSC–derived cardiomyocytes (CMs) carrying the PLN R14del mutation and their corresponding isogenic controls in 3-dimensional engineered heart tissues (EHTs; 2 batches, n=5–12 EHTs each). F and G, Two-dimensional 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. Data are presented as mean±SEM. HDR indicates homology-directed repair. *P<0.0005.
RkJQdWJsaXNoZXIy MTk4NDMw