Figure 4.

Time-Course Study of SNP-Selective Knockdown

(A) mRNA knockdown in hiPSC-derived cardiomyocytes was evaluated at six time points over a 2-week period using allele-specific droplet digital PCR. At day 0, 250 nM rs715-T targeting ASO A259 was added via gymnosis. SNP-matched knockdown is seen for up to 2 weeks, whereas the SNP-mismatched allele does not show knockdown. Data were normalized to the no ASO day 2 time point for each allele. Mean ± SD from three independent experiments is shown. Significance between T alleles (no ASO versus ASO) was determined by two-way ANOVA followed by Sidak’s multiple comparisons test (*p < 0.05, ***p < 0.001). (B) Quantification of β-MHC in iCell² human iPSC-derived cardiomyocyte (hiPSC-CM) with and without addition of ASO A259. β-MHC is not reduced at any time point, suggesting protein compensation by the SNP-mismatched allele. Bar graph represents three independent experiments (mean ± SD). Protein levels were normalized to the no ASO group at each time point.