Live attenuated pseudorabies virus developed using the CRISPR/Cas9 system.
Currently, pseudorabies virus (PRV) variant strains are outbreaking in China; these variants belong to genotype II PRV. The traditional Bartha-K61 vaccine has failed to provide complete protection against the emergent variant strains. Therefore, rapid attenuation of current epidemic strains is needed for effective PRV control. In this study, we report a rapid method for editing the PRV genome using the CRISPR-Cas9 system. We developed a triple gE/gI/TK gene-inactivated HeN1 PRV strain, because mice were more susceptible to PRV infection, we then evaluated the attenuation of PRV in the mice and demonstrated that modified PRV was fully attenuated. Furthermore, the attenuated strain also induced immune protection in response to a parental PRV challenge. Overall, we showed that PRVs can be rapidly attenuated using CRISPR-Cas9 technology, which will be critical for PRV control, especially when new variant PRV strains emerge.
State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin 150001, China.
Yan-Dong
YD
Tang
State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin 150001, China; College of Animal Science and Technology, Jilin Agriculture University, Changchun 130018, China.
Ji-Ting
JT
Liu
State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin 150001, China.
Tong-Yun
TY
Wang
State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin 150001, China.
Tong-Qing
TQ
An
State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin 150001, China.
Ming-Xia
MX
Sun
State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin 150001, China.
Shu-Jie
SJ
Wang
State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin 150001, China.
Qiong-Qiong
QQ
Fang
State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin 150001, China.
Lin-Lin
LL
Hou
State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin 150001, China.
Zhi-Jun
ZJ
Tian
State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin 150001, China. Electronic address: cai139@hvri.ac.cn.
Xue-Hui
XH
Cai
D000818:Animals; D064113:CRISPR-Cas Systems; D002522:Chlorocebus aethiops; D005260:Female; D000072669:Gene Editing; D018390:Gene Targeting; D005822:Genetic Vectors / Q000235:genetics*; D011558:Herpesvirus 1, Suid / Q000235:genetics* / Q000276:immunology; D051379:Mice; D022421:Pseudorabies Vaccines / Q000235:genetics* / Q000276:immunology; D000094704:RNA, Guide, CRISPR-Cas Systems; D017384:Sequence Deletion; D014613:Vaccines, Attenuated / Q000235:genetics* / Q000276:immunology; D014709:Vero Cells; D014779:Virus Replication
D022421:Pseudorabies Vaccines; D000094704:RNA, Guide, CRISPR-Cas Systems; D014613:Vaccines, Attenuated
Attenuation; CRISPR/Cas9; Pseudorabies virus; Vaccine