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

• Published in: Applied microbiology and biotechnology 2024-12, Vol.108 (1), p.248-248, Article 248
• Main Authors: Li, Fengsai, Zhao, Haiyuan, Sui, Ling, Yin, Fangjie, Liu, Xinzi, Guo, Guihai, Li, Jiaxuan, Jiang, Yanping, Cui, Wen, Shan, Zhifu, Zhou, Han, Wang, Li, Qiao, Xinyuan, Tang, Lijie, Wang, Xiaona, Li, Yijing
• Format: Article
• Language: English
• Subjects: Alanine; Alanine racemase; Animals; Anti-Bacterial Agents; Antibiotics; Antibodies, Viral; Applied Genetics and Molecular Biotechnology; Bacteria; Biomedical and Life Sciences; Biotechnology; Clustered Regularly Interspaced Short Palindromic Repeats; CRISPR; Diarrhea; Drug resistance; Economic impact; Epidemics; Gene expression; Genes; Genome editing; Genomes; Homologous recombination; Immune response; Immune response (cell-mediated); Immune response (humoral); Immune system; Immunization; Immunogenicity; Infectious diseases; Lactic acid; Lactic acid bacteria; Life Sciences; Mice; Microbial Genetics and Genomics; Microbiology; Mucosal immunity; Oral administration; Plasmids; Porcine epidemic diarrhea virus - genetics; S1 gene; Swine; Thymidylate synthase; Transmissible gastroenteritis; Transposition; Vaccines; Viral Vaccines; Viruses
• ISSN: 0175-7598; 1432-0614
• DOI: 10.1007/s00253-023-12989-0

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