title: Pseudorabies virus can escape from CRISPR-Cas9-mediated inhibition.
abstract: The CRISPR-Cas9 system is a newly developed genome-engineering tool used to inhibit virus infection by targeting the conserved regions of the viral genomic DNA. In the present study, we constructed a cell line stably expressing Cas9 endonuclease and sgRNA targeting the conserved UL30 gene of pseudorabies virus (PRV). During the PRV infection, the CRISPR-Cas9 system was efficient in cleaving the UL30 gene in each passage. However, deletions and insertions occurred at low passages, while substitutions were frequently observed at high passages. Furthermore, copy numbers and virus titers of PRV were significantly increased in a passage-dependent manner, indicating that viral genomic replication and assembly were more effective at the high passages than at low passages. These results demonstrated that PRV could escape from CRISPR-Cas9-mediated inhibition. Therefore, whether the CRISPR-Cas9 system is suitable for antiviral application should be considered and carefully verified.
authors: Jilin Provincial Key Laboratory of Animal Embryo Engineering, College of Animal Sciences, Jilin University, Changchun, Jilin, 130062, PR China.
authors: Zhiyuan
authors: Z
authors: Peng
authors: Jilin Provincial Key Laboratory of Animal Embryo Engineering, College of Animal Sciences, Jilin University, Changchun, Jilin, 130062, PR China.
authors: Ting
authors: T
authors: Ouyang
authors: Jilin Provincial Key Laboratory of Animal Embryo Engineering, College of Animal Sciences, Jilin University, Changchun, Jilin, 130062, PR China.
authors: Daxin
authors: D
authors: Pang
authors: Jilin Provincial Key Laboratory of Animal Embryo Engineering, College of Animal Sciences, Jilin University, Changchun, Jilin, 130062, PR China.
authors: Teng
authors: T
authors: Ma
authors: Jilin Provincial Key Laboratory of Animal Embryo Engineering, College of Animal Sciences, Jilin University, Changchun, Jilin, 130062, PR China.
authors: Xinrong
authors: X
authors: Chen
authors: Jilin Provincial Key Laboratory of Animal Embryo Engineering, College of Animal Sciences, Jilin University, Changchun, Jilin, 130062, PR China.
authors: Ning
authors: N
authors: Guo
authors: Jilin Provincial Key Laboratory of Animal Embryo Engineering, College of Animal Sciences, Jilin University, Changchun, Jilin, 130062, PR China.
authors: Fuwang
authors: F
authors: Chen
authors: Jilin Provincial Key Laboratory of Animal Embryo Engineering, College of Animal Sciences, Jilin University, Changchun, Jilin, 130062, PR China.
authors: Lin
authors: L
authors: Yuan
authors: Jilin Provincial Key Laboratory of Animal Embryo Engineering, College of Animal Sciences, Jilin University, Changchun, Jilin, 130062, PR China.
authors: Hongsheng
authors: H
authors: Ouyang
authors: Jilin Provincial Key Laboratory of Animal Embryo Engineering, College of Animal Sciences, Jilin University, Changchun, Jilin, 130062, PR China. Electronic address: renlz@jlu.edu.cn.
authors: Linzhu
authors: L
authors: Ren
mesh_terms: D001483:Base Sequence; D064113:CRISPR-Cas Systems; D002460:Cell Line; D004261:DNA Replication; D004279:DNA, Viral; D015870:Gene Expression; D023061:Gene Order; D018390:Gene Targeting; D005822:Genetic Vectors / Q000235:genetics; D011558:Herpesvirus 1, Suid / Q000235:genetics*; D000094704:RNA, Guide, CRISPR-Cas Systems; D014779:Virus Replication / Q000235:genetics*
chemical_list: D004279:DNA, Viral; D000094704:RNA, Guide, CRISPR-Cas Systems
keywords: CRISPR/Cas9; Escape; Inhibition; Pseudorabies virus (PRV); Single-guide RNA (sgRNA); UL30 protein