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Int J Biol Sci 2019; 15(2):481-492. doi:10.7150/ijbs.25862

Research Paper

Generation of Pigs Resistant to Highly Pathogenic-Porcine Reproductive and Respiratory Syndrome Virus through Gene Editing of CD163

Jingyao Chen1, Haitao Wang1, Jianhui Bai1, Wenjie Liu1, Xiaojuan Liu1, Dawei Yu1, Tao Feng2, Zhaolin Sun1, Linlin Zhang1, Linyuan Ma1, Yiqing Hu1, Yunlong Zou1, Tan Tan1, Jie Zhong1, Man Hu1, Xiaofei Bai4, Dengke Pan3, Yiming Xing1, Yaofeng Zhao1, Kegong Tian4,5✉, Xiaoxiang Hu1✉, Ning Li1✉

1. State Key Laboratory for Agrobiotechnology, China Agricultural University, Beijing, China
2. College of animal science and technology, China Agricultural University, Beijing, China
3. Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing, China
4. National Research Center for Veterinary Medicine, Luoyang, Henan Province, China
5. College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou, Henan Province, China

Abstract

Porcine reproductive and respiratory syndrome (PRRS) is a highly contagious disease and the most economically important disease of the swine industry worldwide. Highly pathogenic-PRRS virus (HP-PRRSV) is a variant of PRRSV, which caused high morbidity and mortality. Scavenger receptor CD163, which contains nine scavenger receptor cysteine-rich (SRCR) domains, is a key entry mediator for PRRSV. A previous study demonstrated that SRCR domain 5 (SRCR5), encoded by exon 7, was essential for PRRSV infection in vitro. Here, we substituted exon 7 of porcine CD163 with the corresponding exon of human CD163-like 1 (hCD163L1) using a CRISPR/Cas9 system combined with a donor vector. In CD163Mut/Mut pigs, modifying CD163 gene had no adverse effects on hemoglobin-haptoglobin (Hb-Hp) complex clearance or erythroblast growth. In vitro infection experiments showed that the CD163 mutant strongly inhibited HP-PRRSV replication by inhibiting virus uncoating and genome release. Compared to wild-type (WT) pigs in vivo, HP-PRRSV-infected CD163Mut/Mut pigs showed a substantially decreased viral load in blood and relief from PRRSV-induced fever. While all WT pigs were dead, there of four CD163Mut/Mut pigs survived and recovered at the termination of the experiment. Our data demonstrated that modifying CD163 remarkably inhibited PRRSV replication and protected pigs from HP-PRRSV infection, thus establishing a good foundation for breeding PRRSV-resistant pigs via gene editing technology.

Keywords: HP-PRRSV, CD163, CRISPR/Cas9

This is an open access article distributed under the terms of the Creative Commons Attribution (CC BY-NC) license (https://creativecommons.org/licenses/by-nc/4.0/). See http://ivyspring.com/terms for full terms and conditions.
How to cite this article:
Chen J, Wang H, Bai J, Liu W, Liu X, Yu D, Feng T, Sun Z, Zhang L, Ma L, Hu Y, Zou Y, Tan T, Zhong J, Hu M, Bai X, Pan D, Xing Y, Zhao Y, Tian K, Hu X, Li N. Generation of Pigs Resistant to Highly Pathogenic-Porcine Reproductive and Respiratory Syndrome Virus through Gene Editing of CD163. Int J Biol Sci 2019; 15(2):481-492. doi:10.7150/ijbs.25862. Available from http://www.ijbs.com/v15p0481.htm