An efficient gene-editing technique for use in human pluripotent stem cells (hPSCs) has great potential value in regenerative medicine, as well as in drug discovery based on isogenic human disease models. However, the extremely low efficiency of gene editing in hPSCs remains as a major technical hurdle. Previously, we demonstrated that YM155, a survivin inhibitor developed as an anti-cancer drug, induces highly selective cell death in undifferentiated hPSCs. In this study, we demonstrated that the high cytotoxicity of YM155 in hPSCs, which is mediated by selective cellular uptake of the drug, is due to the high expression of SLC35F2 in these cells. Knockout of SLC35F2 with CRISPR-Cas9, or depletion with siRNAs, made the hPSCs highly resistant to YM155. Simultaneous editing of a gene of interest and transient knockdown of SLC35F2 following YM155 treatment enabled the survival of genome-edited hPSCs as a result of temporary YM155 resistance, thereby achieving an enriched selection of clonal populations with gene knockout or knock-in. This precise and efficient genome editing approach took as little as 3 weeks and required no cell sorting or the introduction of additional genes, to be a more feasible approach for gene editing in hPSCs due to its simplicity.

用于人类多能干细胞（hPSC）的有效基因编辑技术在再生医学以及基于等基因人类疾病模型的药物发现中具有巨大的潜在价值。但是，hPSCs中基因编辑的极低效率仍然是主要的技术障碍。以前，我们证明YM155是一种作为抗癌药开发的生存素抑制剂，可在未分化的hPSC中诱导高度选择性的细胞死亡。在这项研究中，我们证明了YM155在hPSCs中的高细胞毒性是由SLC35F2的高表达引起的，这是由药物的选择性细胞摄取介导的在这些细胞中。用CRISPR-Cas9敲除SLC35F2或siRNA耗尽，使得hPSC对YM155具有高度抗性。YM155处理后，对目标基因的同时编辑和SLC35F2的瞬时敲除可实现基因组编辑的hPSC的存活，这是暂时的YM155抗性的结果，从而通过基因敲除或敲入获得了丰富的克隆种群选择。这种精确而有效的基因组编辑方法耗时仅3周，并且不需要细胞分选或引入其他基因，由于其简单性，使其成为hPS​​C中进行基因编辑的更可行方法。