AUTHOR=Zhao Heng , Ye Weijian , Guo Jianxiong , Wang Jiaoxiang , Jiao Deling , Xu Kaixiang , Yang Chang , Chen Shuhan , Jamal Muhammad Ameen , Bai Zhongbin , Wei Taiyun , Cai Jie , Nguyen Tien Dat , Qing Yubo , Cheng Wenmin , Jia Baoyu , Li Honghui , Zhao Hong-Ye , Chen Qingfeng , Wei Hong-Jiang 

TITLE=Development of RAG2-/-IL2Rγ-/Y immune deficient FAH-knockout miniature pig

JOURNAL=Frontiers in Immunology

VOLUME=13

YEAR=2022

URL=https://www.frontiersin.org/journals/immunology/articles/10.3389/fimmu.2022.950194

DOI=10.3389/fimmu.2022.950194

ISSN=1664-3224

ABSTRACT=<p>Human hepatocyte transplantation for liver disease treatment have been hampered by the lack of quality human hepatocytes. Pigs with their large body size, longevity and physiological similarities with human are appropriate animal models for the <italic>in vivo</italic> expansion of human hepatocytes. Here we report on the generation of RAG2<sup>-/-</sup>IL2Rγ<sup>-/Y</sup>FAH<sup>-/-</sup> (RGFKO) pigs <italic>via</italic> CRISPR/Cas9 system and somatic cell nuclear transfer. We showed that thymic and splenic development in RGFKO pigs was impaired. V(D)J recombination processes were also inactivated. Consequently, RGFKO pigs had significantly reduced numbers of porcine T, B and NK cells. Moreover, due to the loss of FAH, porcine hepatocytes continuously undergo apoptosis and consequently suffer hepatic damage. Thus, RGFKO pigs are both immune deficient and constantly suffer liver injury in the absence of NTBC supplementation. These results suggest that RGFKO pigs have the potential to be engrafted with human hepatocytes without immune rejection, thereby allowing for large scale expansion of human hepatocytes.</p>