{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,22]],"date-time":"2026-10-22T14:17:53Z","timestamp":1792678673939,"version":"4.5.0"},"reference-count":50,"publisher":"American Society for Microbiology","issue":"12","license":[{"start":{"date-parts":[[2023,12,21]],"date-time":"2023-12-21T00:00:00Z","timestamp":1703116800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/doi.org\/10.1128\/ASMCopyrightv2"},{"start":{"date-parts":[[2023,12,21]],"date-time":"2023-12-21T00:00:00Z","timestamp":1703116800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/100000199","name":"U.S. Department of Agriculture","doi-asserted-by":"crossref","award":["2019-67015-29843"],"award-info":[{"award-number":["2019-67015-29843"]}],"id":[{"id":"10.13039\/100000199","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["J Virol"],"published-print":{"date-parts":[[2023,12,21]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            Porcine epidemic diarrhea virus (PEDV) is a deadly coronavirus for neonatal piglets and no effective vaccines are available. Transcriptional regulatory sequences (TRSs) are critical in regulating coronavirus discontinuous transcription. Also, TRSs contribute to a high recombination rate of coronaviruses, leading to difficulty in developing safe live vaccines. We hypothesize that recoding the TRS core sequences (TRS-CSs) of PEDV can make the recombination impossible between the engineered vaccine virus and field strains or wild-type viruses. We used an infectious clone-derived reporter PEDV, dORF3-enhanced green fluorescent protein (EGFP), as the backbone to generate a remodeled TRS (RMT) mutant that carries the recoded leader and body TRS-CSs (except for the EGFP). The RMT and dORF3-EGFP showed comparable replication efficiency in Vero cells. However, the incompatibility between the rewired and wild-type TRS-CSs led to few EGFP in RMT-infected cells. Furthermore, RMT and dORF3-EGFP had a similar attenuated phenotype, replication efficiency, and protective immunogenicity in neonatal pigs. RNA sequencing analysis indicated that EGFP transcription directed by the heterogeneous TRS-CSs was significantly reduced to an extremely low level. Meanwhile, recombinant viruses were not detected in Vero cells and in pigs that were co-infected with RMT and a PEDV S-INDEL strain, Iowa106.\n            <jats:italic>In vitro<\/jats:italic>\n            and\n            <jats:italic>in vivo<\/jats:italic>\n            passaging of the RMT did not result in reversion mutations in the rewired TRS-CSs, introduced gaps, and disrupted wild-type TRSs. In summary, the RMT mutant was resistant to recombination and genetically stable and can be further optimized (e.g., deletion of the EGFP) to serve as a platform to develop safe PEDV live attenuated vaccines.\n          <\/jats:p>\n          <jats:sec>\n            <jats:title>IMPORTANCE<\/jats:title>\n            <jats:p>Coronaviruses are important pathogens of humans and animals, and vaccine developments against them are imperative. Due to the ability to induce broad and prolonged protective immunity and the convenient administration routes, live attenuated vaccines (LAVs) are promising arms for controlling the deadly coronavirus infections. However, potential recombination events between vaccine and field strains raise a safety concern for LAVs. The porcine epidemic diarrhea virus (PEDV) remodeled TRS (RMT) mutant generated in this study replicated efficiently in both cell culture and in pigs and retained protective immunogenicity against PEDV challenge in pigs. Furthermore, the RMT PEDV was resistant to recombination and genetically stable. Therefore, RMT PEDV can be further optimized as a backbone for the development of safe LAVs.<\/jats:p>\n          <\/jats:sec>","DOI":"10.1128\/jvi.01193-23","type":"journal-article","created":{"date-parts":[[2023,11,16]],"date-time":"2023-11-16T14:01:59Z","timestamp":1700143319000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":17,"title":["A recombination-resistant genome for live attenuated and stable PEDV vaccines by engineering the transcriptional regulatory sequences"],"prefix":"10.1128","volume":"97","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6412-7348","authenticated-orcid":false,"given":"Xiaoyu","family":"Niu","sequence":"first","affiliation":[{"name":"Center for Food Animal Health, Department of Animal Sciences, College of Food, Agricultural and Environmental Sciences, The Ohio State University, Wooster, Ohio, USA"},{"name":"Department of Veterinary Preventive Medicine, College of Veterinary Medicine, The Ohio State University, Columbus, Ohio, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mingde","family":"Liu","sequence":"additional","affiliation":[{"name":"Center for Food Animal Health, Department of Animal Sciences, College of Food, Agricultural and Environmental Sciences, The Ohio State University, Wooster, Ohio, USA"},{"name":"Department of Veterinary Preventive Medicine, College of Veterinary Medicine, The Ohio State University, Columbus, Ohio, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7932-4777","authenticated-orcid":false,"given":"Shaomin","family":"Yang","sequence":"additional","affiliation":[{"name":"Department of Pain Medicine, Shenzhen Nanshan People\u2019s Hospital and the 6th Affiliated Hospital of Guangdong Medical University, Shenzhen, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiayu","family":"Xu","sequence":"additional","affiliation":[{"name":"Department of Veterinary Preventive Medicine, College of Veterinary Medicine, The Ohio State University, Columbus, Ohio, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yixuan J.","family":"Hou","sequence":"additional","affiliation":[{"name":"Center for Food Animal Health, Department of Animal Sciences, College of Food, Agricultural and Environmental Sciences, The Ohio State University, Wooster, Ohio, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dongxiao","family":"Liu","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Howard University College of Medicine, Washington, DC, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6487-2356","authenticated-orcid":false,"given":"Qiyi","family":"Tang","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Howard University College of Medicine, Washington, DC, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6087-3175","authenticated-orcid":false,"given":"Hua","family":"Zhu","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Molecular Genetics, UMDNJ-New Jersey Medical School, Newark, New Jersey, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8238-417X","authenticated-orcid":false,"given":"Qiuhong","family":"Wang","sequence":"additional","affiliation":[{"name":"Center for Food Animal Health, Department of Animal Sciences, College of Food, Agricultural and Environmental Sciences, The Ohio State University, Wooster, Ohio, USA"},{"name":"Department of Veterinary Preventive Medicine, College of Veterinary Medicine, The Ohio State University, Columbus, Ohio, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1136\/vr.100.12.243"},{"key":"e_1_3_3_3_2","doi-asserted-by":"publisher","DOI":"10.1007\/BF01317606"},{"key":"e_1_3_3_4_2","doi-asserted-by":"publisher","DOI":"10.3201\/eid1803.120002"},{"key":"e_1_3_3_5_2","doi-asserted-by":"publisher","DOI":"10.3201\/eid1801.111259"},{"key":"e_1_3_3_6_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.virusres.2016.05.026"},{"key":"e_1_3_3_7_2","first-page":"1320","article-title":"Characterization of the structural proteins of porcine epizootoc diarrhea virus, strain CV777","volume":"49","author":"Egberink HF","year":"1988","unstructured":"Egberink HF, Ederveen J, Callebaut P, Horzinek MC. 1988. 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