Generation of complement protein C3 deficient pigs by CRISPR/Cas9-mediated gene targeting

Scientific Reports

卷:7

Wei Zhang¹  Yong Jin¹  Lihua Zhao¹  Lining Zhang¹  Rongfeng Li¹  Guan Wang¹  Qiang Xiong¹  Haiyuan Yang¹  Ying Wang¹  Yifan Dai¹  Lisha Mou² 

[1] Jiangsu Key Laboratory of Xenotransplantation, Nanjing Medical University;

[2] Shenzhen Xenotransplantation Medical Engineering Research and Development Center, Institute of Translational Medicine, Shenzhen Second People’s Hospital, First Affiliated Hospital of Shenzhen University;

关键词: ;   

DOI  :  10.1038/s41598-017-05400-2

来源: DOAJ

【 摘 要 】

Abstract Complement protein C3 is the pivotal component of the complement system. Previous studies have demonstrated that C3 has implications in various human diseases and exerts profound functions under certain conditions. However, the delineation of pathological and physiological roles of C3 has been hampered by the insufficiency of suitable animal models. In the present study, we applied the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated (Cas) system to target the C3 gene in porcine fetal fibroblasts. Our results indicated that CRISPR/Cas9 targeting efficiency was as high as 84.7%, and the biallelic mutation efficiency reached at 45.7%. The biallelic modified colonies were used as donor for somatic cell nuclear transfer (SCNT) technology to generate C3 targeted piglets. A total of 19 C3 knockout (KO) piglets were produced and their plasma C3 protein was undetectable by western blot analysis and ELISA. The hemolytic complement activity and complement-dependent cytotoxicity assay further confirmed that C3 was disrupted in these piglets. These C3 KO pigs could be utilized as a valuable large animal model for the elucidation of the roles of C3.

【 授权许可】

Unknown