The clinical applicability of porcine xenotransplantation—a long-investigated alternative to the scarce availability of human organs for patients with organ failure—is limited by molecular incompatibilities between the immune systems of pigs and humans as well as by the risk of transmitting porcine endogenous retroviruses (PERVs). We recently showed the production of pigs with genomically inactivated PERVs. Here, using a combination of CRISPR–Cas9 and transposon technologies, we show that pigs with all PERVs inactivated can also be genetically engineered to eliminate three xenoantigens and to express nine human transgenes that enhance the pigs’ immunological compatibility and blood-coagulation compatibility with humans. The engineered pigs exhibit normal physiology, fertility and germline transmission of the 13 genes and 42 alleles edited. Using in vitro assays, we show that cells from the engineered pigs are resistant to human humoral rejection, cell-mediated damage and pathogenesis associated with dysregulated coagulation. The extensive genome engineering of pigs for greater compatibility with the human immune system may eventually enable safe and effective porcine xenotransplantation.

猪异种移植的临床适用性——一种长期研究的替代人体器官的稀缺可用于器官衰竭患者——受到猪和人类免疫系统之间的分子不相容性以及传播猪内源性逆转录病毒的风险的限制。 PERV)。我们最近展示了具有基因组灭活 PERV 的猪的生产情况。在这里，我们结合使用 CRISPR–Cas9 和转座子技术，表明所有 PERV 失活的猪也可以通过基因工程消除三种异种抗原并表达九种人类转基因，从而增强猪与人类的免疫相容性和凝血相容性. 工程猪表现出正常的生理机能、生育能力和编辑的 13 个基因和 42 个等位基因的种系传递。使用体外试验，我们表明来自工程猪的细胞对人体体液排斥、细胞介导的损伤和与凝血失调相关的发病机制具有抵抗力。猪的广泛基因组工程与人类免疫系统具有更大的相容性，最终可能实现安全有效的猪异种移植。