{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,16]],"date-time":"2026-04-16T08:18:22Z","timestamp":1776327502087,"version":"3.50.1"},"reference-count":30,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2023,10,10]],"date-time":"2023-10-10T00:00:00Z","timestamp":1696896000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Microbiol."],"abstract":"<jats:p>Since 2011, pseudorabies based on the pseudorabies virus (PRV) variant has emerged as a serious health issue in pig farms in China. The PRV gE\/TK or gE\/gI\/TK deletion strains protect against emerging PRV variants. However, these variants may cause lethal infections in newborn piglets without PRV antibodies. Previous studies have shown that codon deoptimization of a virulence gene causes virus attenuation. Accordingly, we deoptimized US3-S (US3 gene encoding a short isoform that represents approximately 95% of the total US3 transcription) and <jats:italic>UL56<\/jats:italic> genes (first 10 or all codons) of PRV gE\/TK deletion strain (PRV<jats:sup>\u0394TK&amp;amp;gE\u2212AH02<\/jats:sup>) to generate six recombinant PRVs through bacterial artificial chromosome technology. In swine testicular cells, recombinant PRVs with all codon deoptimization of <jats:italic>US3-S<\/jats:italic> or <jats:italic>UL56<\/jats:italic> genes were grown to lower titers than the parental virus. Notably, US3-S or UL56 with all codon deoptimization reduced mRNA and protein expressions. Subsequently, the safety and immunogenicity of recombinant PRVs with codon deoptimization of US3-S or UL56 are evaluated as vaccine candidates in mice and piglets. The mice inoculated with recombinant PRVs with codon deoptimization of US3-S or UL56 showed exceptional survival ability without severe clinical signs. All codons deoptimized (US3-S and UL56) significantly decreased virus load and attenuated pathological changes in the brains of the mice. Moreover, the protection efficiency offered by recombinant PRVs with codon deoptimization of US3-S or UL56 showed similar effects to PRV<jats:sup>\u0394TK&amp;amp;gE\u2212AH02<\/jats:sup>. Remarkably, the 1-day-old PRV antibody-negative piglets inoculated with PRV<jats:sup>\u0394TK&amp;amp;gE<\/jats:sup>-US3-S<jats:sup>T\u2212CD<\/jats:sup> (a recombinant PRV with all codon deoptimization of US3-S) presented no abnormal clinical symptoms, including fever. The piglets inoculated with PRV<jats:sup>\u0394TK&amp;amp;gE<\/jats:sup>-US3-S<jats:sup>T\u2212CD<\/jats:sup> showed a high serum neutralization index against the PRV variant. In conclusion, these results suggest using codon deoptimization to generate innovative live attenuated PRV vaccine candidates.<\/jats:p>","DOI":"10.3389\/fmicb.2023.1248573","type":"journal-article","created":{"date-parts":[[2023,10,10]],"date-time":"2023-10-10T05:27:18Z","timestamp":1696915638000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":4,"title":["Construction of pseudorabies virus variant attenuated vaccine: codon deoptimization of US3 and UL56 genes based on PRV gE\/TK deletion strain"],"prefix":"10.3389","volume":"14","author":[{"given":"Mengwei","family":"Xu","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Laixu","family":"Zhu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Aimin","family":"Ge","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yamei","family":"Liu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Saisai","family":"Chen","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ziwen","family":"Wei","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yating","family":"Zheng","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ling","family":"Tong","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zhisheng","family":"Wang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rongmei","family":"Fei","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jichun","family":"Wang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chuanjian","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1965","published-online":{"date-parts":[[2023,10,10]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"1749","DOI":"10.3201\/eid1911.130177","article-title":"Pseudorabies virus variant in Bartha-K61-vaccinated pigs, China, 2012","volume":"19","author":"An","year":"2013","journal-title":"Emerg. Infect. Dis."},{"key":"B2","doi-asserted-by":"publisher","first-page":"3259","DOI":"10.1128\/JVI.80.7.3259-3272.2006","article-title":"Modulation of poliovirus replicative fitness in HeLa cells by deoptimization of synonymous codon usage in the capsid region","volume":"80","author":"Burns","year":"2006","journal-title":"J. Virol."},{"key":"B3","doi-asserted-by":"publisher","first-page":"e00039","DOI":"10.1128\/mBio.00039-20","article-title":"A Lassa fever live-attenuated vaccine based on codon deoptimization of the viral glycoprotein gene","volume":"11","author":"Cai","year":"2020","journal-title":"mBio"},{"key":"B4","doi-asserted-by":"publisher","first-page":"156","DOI":"10.1016\/0003-2697(87)90021-2","article-title":"Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction","volume":"162","author":"Chomczynski","year":"1987","journal-title":"Anal. Biochem."},{"key":"B5","doi-asserted-by":"publisher","first-page":"179","DOI":"10.1016\/j.virol.2015.11.014","article-title":"The pseudorabies virus protein, pUL56, enhances virus dissemination and virulence but is dispensable for axonal transport","volume":"488","author":"Daniel","year":"2016","journal-title":"Virology"},{"key":"B6","doi-asserted-by":"publisher","first-page":"1298","DOI":"10.1128\/JVI.02167-15","article-title":"Synonymous deoptimization of foot-and-mouth disease virus causes attenuation in vivo while inducing a strong neutralizing antibody response","volume":"90","author":"Diaz-San Segundo","year":"2016","journal-title":"J. Virol."},{"key":"B7","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1006857","article-title":"Attenuation of a very virulent Marek's disease herpesvirus (MDV) by codon pair bias deoptimization","author":"Eschke","year":"2018","journal-title":"PLoS Pathog"},{"key":"B8","doi-asserted-by":"publisher","first-page":"3","DOI":"10.1016\/j.vetmic.2016.11.019","article-title":"Vaccines against pseudorabies virus (PrV)","volume":"206","author":"Freuling","year":"2017","journal-title":"Vet. Microbiol."},{"key":"B9","doi-asserted-by":"publisher","first-page":"e02238","DOI":"10.1128\/mBio.02238-20","article-title":"Mechanisms of attenuation by genetic recoding of viruses","volume":"12","author":"Goncalves-Carneiro","year":"2021","journal-title":"mBio"},{"key":"B10","doi-asserted-by":"publisher","first-page":"290","DOI":"10.1007\/s002390010219","article-title":"Codon usage and tRNA genes in eukaryotes: correlation of codon usage diversity with translation efficiency and with CG-dinucleotide usage as assessed by multivariate analysis","volume":"53","author":"Kanaya","year":"2001","journal-title":"J. Mol. Evol."},{"key":"B11","doi-asserted-by":"publisher","first-page":"511","DOI":"10.1006\/viro.1994.1672","article-title":"Inactivation of glycoprotein gE and thymidine kinase or the US3-encoded protein kinase synergistically decreases in vivo replication of pseudorabies virus and the induction of protective immunity","volume":"205","author":"Kimman","year":"1994","journal-title":"Virology"},{"key":"B12","doi-asserted-by":"publisher","DOI":"10.1186\/gb-2001-2-4-research0010","article-title":"A simple model based on mutation and selection explains trends in codon and amino-acid usage and GC composition within and across genomes","author":"Knight","year":"2001","journal-title":"Genome Biol."},{"key":"B13","doi-asserted-by":"publisher","first-page":"1708","DOI":"10.1016\/j.vaccine.2021.02.024","article-title":"Vaccine candidates generated by codon and codon pair deoptimization of enterovirus A71 protect against lethal challenge in mice","volume":"39","author":"Lee","year":"2021","journal-title":"Vaccine"},{"key":"B14","doi-asserted-by":"publisher","first-page":"e0027222","DOI":"10.1128\/spectrum.00272-22","article-title":"Vaccinia virus attenuation by codon deoptimization of the a24r gene for vaccine development","volume":"10","author":"Lorenzo","year":"2022","journal-title":"Microbiol. Spectr."},{"key":"B15","first-page":"95","article-title":"Construction and biological characterization of pseudorabies virus variant of the inactivated US3 gene","volume":"52","author":"Lv","year":"","journal-title":"Anim. Husb. Vet. Med."},{"key":"B16","first-page":"34","article-title":"Construction of UL43 and UL56 gene inactivation mutants of pseudorabies virus variant and analysis of their biological characteristics","volume":"30","author":"Lv","year":"","journal-title":"Chin. J. Anim. Infect. Dis."},{"key":"B17","doi-asserted-by":"publisher","first-page":"e01704","DOI":"10.1128\/mBio.01704-14","article-title":"Refining the balance of attenuation and immunogenicity of respiratory syncytial virus by targeted codon deoptimization of virulence genes","volume":"5","author":"Meng","year":"2014","journal-title":"mBio"},{"key":"B18","doi-asserted-by":"publisher","first-page":"10525","DOI":"10.1128\/JVI.01565-14","article-title":"Influenza A virus attenuation by codon deoptimization of the NS gene for vaccine development","volume":"88","author":"Nogales","year":"2014","journal-title":"J. Virol."},{"key":"B19","doi-asserted-by":"publisher","first-page":"6387","DOI":"10.1128\/JVI.00352-06","article-title":"Role of pseudorabies virus Us3 protein kinase during neuronal infection","volume":"80","author":"Olsen","year":"2006","journal-title":"J. Virol."},{"key":"B20","doi-asserted-by":"publisher","first-page":"462","DOI":"10.1128\/MMBR.69.3.462-500.2005","article-title":"Molecular biology of pseudorabies virus: impact on neurovirology and veterinary medicine","volume":"69","author":"Pomeranz","year":"2005","journal-title":"Microbiol. Mol. Biol. Rev."},{"key":"B21","doi-asserted-by":"publisher","first-page":"e02029","DOI":"10.1128\/JVI.02029-19","article-title":"Roles of the different isoforms of the pseudorabies virus protein kinase pUS3 in nuclear egress","volume":"94","author":"Sehl","year":"2020","journal-title":"J. Virol."},{"key":"B22","doi-asserted-by":"publisher","first-page":"421","DOI":"10.1007\/978-1-60761-652-8_30","article-title":"En passant mutagenesis: a two step markerless red recombination system","volume":"634","author":"Tischer","year":"2010","journal-title":"Methods Mol. Biol."},{"key":"B23","doi-asserted-by":"publisher","first-page":"287","DOI":"10.1186\/s12917-018-1536-7","article-title":"Safety and immunogenicity of an attenuated Chinese pseudorabies variant by dual deletion of TKandgE genes","volume":"14","author":"Wang","year":"2018","journal-title":"BMC. Vet. Res."},{"key":"B24","doi-asserted-by":"publisher","DOI":"10.1128\/jvi.01545-22","article-title":"Pseudorabies virus regulates the extracellular translocation of annexin A2 to promote its proliferation. J","author":"Weng","year":"2023","journal-title":"Virol"},{"key":"B25","doi-asserted-by":"publisher","first-page":"198740","DOI":"10.1016\/j.virusres.2022.198740","article-title":"A noval strategy of deletion in PK gene for construction of a vaccine candidate with exellent safety and complete protection efficiency against high virulent Chinese pseudorabies virus variant","volume":"313","author":"Xu","year":"2022","journal-title":"Virus Res."},{"key":"B26","doi-asserted-by":"publisher","first-page":"102","DOI":"10.3201\/eid2001.130531","article-title":"Pathogenic pseudorabies virus, China, 2012","volume":"20","author":"Yu","year":"2014","journal-title":"Emerg. Infect. Dis."},{"key":"B27","doi-asserted-by":"publisher","first-page":"2432","DOI":"10.1016\/j.vaccine.2015.03.094","article-title":"Construction of a triple gene-deleted Chinese Pseudorabies virus variant and its efficacy study as a vaccine candidate on suckling piglets","volume":"33","author":"Zhang","year":"2015","journal-title":"Vaccine"},{"key":"B28","doi-asserted-by":"publisher","first-page":"2","DOI":"10.1186\/s12917-018-1766-8","article-title":"A gDandgC-substituted pseudorabies virus vaccine strain provides complete clinical protection and is helpful to prevent virus shedding against challenge by a Chinese pseudorabies variant","volume":"15","author":"Zhang","year":"","journal-title":"BMC. Vet. Res."},{"key":"B29","doi-asserted-by":"publisher","first-page":"518","DOI":"10.3390\/v11060518","article-title":"Effects of intranasal pseudorabies virus AH02LA infection on microbial community and immune status in the ileum and colon of piglets","volume":"11","author":"Zhang","year":"","journal-title":"Viruses"},{"key":"B30","doi-asserted-by":"publisher","first-page":"1161","DOI":"10.1016\/j.vaccine.2017.01.003","article-title":"Bartha-k61 vaccine protects growing pigs against challenge with an emerging variant pseudorabies virus","volume":"35","author":"Zhou","year":"2017","journal-title":"Vaccine"}],"container-title":["Frontiers in Microbiology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fmicb.2023.1248573\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,10,10]],"date-time":"2023-10-10T05:27:24Z","timestamp":1696915644000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fmicb.2023.1248573\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,10,10]]},"references-count":30,"alternative-id":["10.3389\/fmicb.2023.1248573"],"URL":"https:\/\/doi.org\/10.3389\/fmicb.2023.1248573","relation":{},"ISSN":["1664-302X"],"issn-type":[{"value":"1664-302X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,10,10]]},"article-number":"1248573"}}