Molecular imaging is indispensable for determining the fate and persistence of engrafted stem cells. Standard strategies for transgene induction involve the use of viral vectors prone to silencing and insertional mutagenesis or the use of nonhuman genes. Methods: We used zinc finger nucleases to induce stable expression of human imaging reporter genes into the safe-harbor locus adeno-associated virus integration site 1 in human embryonic stem cells. Plasmids were generated carrying reporter genes for fluorescence, bioluminescence imaging, and human PET reporter genes. Results: In vitro assays confirmed their functionality, and embryonic stem cells retained differentiation capacity. Teratoma formation assays were performed, and tumors were imaged over time with PET and bioluminescence imaging. Conclusion: This study demonstrates the application of genome editing for targeted integration of human imaging reporter genes in human embryonic stem cells for long-term molecular imaging.

分子成像对于确定移植干细胞的命运和持久性是必不可少的。转基因诱导的标准策略包括使用易于沉默和插入诱变的病毒载体或使用非人类基因。方法：我们使用锌指核酸酶诱导人类成像报告基因稳定表达到人类胚胎干细胞中的安全港基因座腺相关病毒整合位点1中。产生带有用于荧光，生物发光成像和人类PET报道基因的报道基因的质粒。结果：体外测定证实了它们的功能，并且胚胎干细胞保留了分化能力。进行畸胎瘤形成测定，并通过PET和生物发光成像对肿瘤随时间成像。结论：这项研究证明了基因组编辑技术在人类胚胎干细胞中有针对性地整合人类成像报告基因以进行长期分子成像的应用。