title: A CRISPR/Cas9 and Cre/Lox system-based express vaccine development strategy against re-emerging Pseudorabies virus.
abstract: Virus evolves rapidly to escape vaccine-induced immunity, posing a desperate demand for efficient vaccine development biotechnologies. Here we present an express vaccine development strategy based on CRISPR/Cas9 and Cre/Lox system against re-emerging Pseudorabies virus, which caused the recent devastating swine pseudorabies outbreak in China. By CRISPR/Cas9 system, the virulent genes of the newly isolated strain were simultaneously substituted by marker genes, which were subsequently excised using Cre/Lox system for vaccine safety concern. Notably, single cell FACS technology was applied to further promote virus purification efficiency. The combination of these state-of-art technologies greatly accelerated vaccine development. Finally, vaccination and challenge experiments proved this vaccine candidate's protective efficacy in pigs and the promise to control current pseudorabies outbreak. This is, to our knowledge, the first successful vaccine development based on gene edit technologies, demonstrating these technologies leap from laboratory to industry. It may pave the way for future express antiviral vaccine development.
authors.affiliation: State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan, 430070, China.|College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, 430070, China.
authors.forename: Xun
authors.initials: X
authors.lastname: Liang
authors.affiliation: State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan, 430070, China.|College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, 430070, China.
authors.forename: Leqiang
authors.initials: L
authors.lastname: Sun
authors.affiliation: State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan, 430070, China.|College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, 430070, China.
authors.forename: Teng
authors.initials: T
authors.lastname: Yu
authors.affiliation: Guangdong Wen's Group Academy, Guangdong Wen's Foodstuffs Group Co.,Ltd., Yunfu, 527300, China.
authors.forename: Yongfei
authors.initials: Y
authors.lastname: Pan
authors.affiliation: Guangdong Wen's Group Academy, Guangdong Wen's Foodstuffs Group Co.,Ltd., Yunfu, 527300, China.
authors.forename: Dongdong
authors.initials: D
authors.lastname: Wang
authors.affiliation: College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, 430070, China.
authors.forename: Xueying
authors.initials: X
authors.lastname: Hu
authors.affiliation: Departments of Pathology, College of Veterinary Medicine, University of Georgia, Athens, GA 30602, USA.
authors.forename: Zhenfang
authors.initials: Z
authors.lastname: Fu
authors.affiliation: State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan, 430070, China.|College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, 430070, China.|Key Laboratory of Development of Veterinary Diagnostic Products, Ministry of Agriculture, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, 430070, China.
authors.forename: Qigai
authors.initials: Q
authors.lastname: He
authors.affiliation: State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan, 430070, China.|College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, 430070, China.|Key Laboratory of Development of Veterinary Diagnostic Products, Ministry of Agriculture, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, 430070, China.
authors.forename: Gang
authors.initials: G
authors.lastname: Cao
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology; D064113:CRISPR-Cas Systems / Q000235:genetics*; D002681:China; D011558:Herpesvirus 1, Suid / Q000187:drug effects / Q000472:pathogenicity; D011557:Pseudorabies / Q000517:prevention & control* / Q000821:virology; D013552:Swine; D013553:Swine Diseases / Q000276:immunology / Q000517:prevention & control*; D014765:Viral Vaccines / Q000276:immunology / Q000627:therapeutic use*
chemical_list: D000914:Antibodies, Viral; D014765:Viral Vaccines