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Yue, Yanan; Xu, Weihong; Kan, Yinan; Zhao, Hong-Ye; Zhou, Yixuan; Song, Xiaobin; Wu, Jiajia; Xiong, Juan; Goswami, Dharmendra; Yang, Meng; Lamriben, Lydia; Xu, Mengyuan; Zhang, Qi; Luo, Yu; Guo, Jianxiong; Mao, Shengyi; Jiao, Deling; Nguyen, Tien Dat; Li, Zhuo; Layer, Jacob V; Li, Mailin; Paragas, Violette; Youd, Michele E; Sun, Zhongquan; Ding, Yuan; Wang, Weilin; Dou, Hongwei; Song, Lingling; Wang, Xueqiong; Le, Lei; Fang, Xin; George, Haydy; Anand, Ranjith; Wang, Shi Yun; Westlin, William F; Güell, Marc; Markmann, James; Qin, Wenning; Gao, Yangbin; Wei, Hong-Jiang; Church, George M; Yang, Luhan
Nature biomedical engineering, 02/2021, Volume: 5, Issue: 2Journal Article
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.
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