Patient-specific human induced pluripotent stem cells (hiPSCs) offer unprecedented opportunities for the investigation of multigenic disease, personalized medicine, and stem cell therapy. For heterogeneous diseases such as atrial fibrillation (AF), however, precise correction of the associated mutation is crucial. Here, we generated and corrected hiPSC lines from two AF patients carrying different heterozygous SHOX2 mutations. We developed a strategy for the scarless correction of heterozygous mutations, based on stochastic enrichment by sib selection, followed by allele quantification via digital PCR and next-generation sequencing to detect isogenic subpopulations. This allowed enriching edited cells 8- to 20-fold. The method does not require antibiotic selection or cell sorting and can be easily combined with base-and-prime editing approaches. Our strategy helps to overcome low efficiencies of homology-dependent repair in hiPSCs and facilitates the generation of isogenic control lines that represent the gold standard for modeling complex diseases in vitro.

患者特异性人类诱导多能干细胞 (hiPSC) 为多基因疾病、个性化医疗和干细胞治疗的研究提供了前所未有的机会。然而，对于心房颤动 (AF) 等异质性疾病，精确纠正相关突变至关重要。在这里，我们从两名携带不同杂合SHOX2突变的 AF 患者中生成并校正了 hiPSC 系。我们开发了一种杂合突变的无痕校正策略，基于同胞选择的随机富集，然后通过数字 PCR 和下一代测序进行等位基因定量以检测同基因亚群。这使得编辑后的细胞富集了 8 至 20 倍。该方法不需要抗生素选择或细胞分选，并且可以轻松地与碱基和引物编辑方法相结合。我们的策略有助于克服 hiPSC 中同源依赖性修复的低效率，并促进同基因对照系的生成，这些对照系代表了体外复杂疾病建模的黄金标准。