Assessing immunogenicity of CRISPR-NCas9 engineered strain against porcine epidemic diarrhea virus

• DOI: 10.1007/s00253-023-12989-0
• Publication Date: 2024-03-02T09:02:08Z
• Authors: Fengsai Li; Haiyuan Zhao; Ling Sui; Fangjie Yin; Xinzi Liu; Guihai Guo; Jiaxuan Li; Yanping Jiang; Wen Cui; Zhifu Shan; Han Zhou; Li Wang; Xinyuan Qiao; Lijie Tang; Xiaona Wang; Yijing Li
• Keywords: Attenuated vaccine; Lactobacillus paracasei

Abstract Porcine epidemic diarrhea (PED) caused by porcine virus (PEDV), is an acute and highly infectious disease, resulting in substantial economic losses the pig industry. Given that PEDV primarily infects mucosal surfaces of intestinal tract, it crucial to improve immunity prevent viral invasion. Lactic acid bacteria (LAB) oral vaccines offer unique advantages potential applications combatting diseases, making them ideal approach for controlling PED outbreaks. However, traditional LAB use plasmids exogenous protein expression antibiotic genes as selection markers. Antibiotic can be diffused through transposition, transfer, or homologous recombination, generation drug-resistant strains. To overcome these issues, genome-editing technology has been developed achieve gene genomes. In this study, we used CRISPR-NCas9 system integrate S1 into genome alanine racemase-deficient Lactobacillus paracasei △ Alr HLJ-27 ( L. ) at thymidylate synthase (thyA) site, generating a strain, S1/ . We conducted immunization assays mice piglets evaluate level immune response evaluated its protective effect against challenge tests piglets. Oral administration strain elicited mucosal, humoral, cellular responses. The also exhibited certain resistance infection These results demonstrate vaccine candidate control. Key points • A S1/△Alr was constructed vaccine. Immunogenicity test carried out analyze ability strain. could provide protection extent.