{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,19]],"date-time":"2026-02-19T05:48:24Z","timestamp":1771480104874,"version":"3.50.1"},"reference-count":36,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2024,2,18]],"date-time":"2024-02-18T00:00:00Z","timestamp":1708214400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Key Research and Development Program of China","award":["2023YFD1802600"],"award-info":[{"award-number":["2023YFD1802600"]}]},{"name":"National Key Research and Development Program of China","award":["2022YFD1800500"],"award-info":[{"award-number":["2022YFD1800500"]}]},{"name":"National Key Research and Development Program of China","award":["2021YFD1801401"],"award-info":[{"award-number":["2021YFD1801401"]}]},{"name":"National Key Research and Development Program of China","award":["Y2022PT11"],"award-info":[{"award-number":["Y2022PT11"]}]},{"name":"Central Public-interest Scientific Institution Basal Research Fund","award":["2023YFD1802600"],"award-info":[{"award-number":["2023YFD1802600"]}]},{"name":"Central Public-interest Scientific Institution Basal Research Fund","award":["2022YFD1800500"],"award-info":[{"award-number":["2022YFD1800500"]}]},{"name":"Central Public-interest Scientific Institution Basal Research Fund","award":["2021YFD1801401"],"award-info":[{"award-number":["2021YFD1801401"]}]},{"name":"Central Public-interest Scientific Institution Basal Research Fund","award":["Y2022PT11"],"award-info":[{"award-number":["Y2022PT11"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>The CRISPR\/Cas9 system is widely used to manipulate viral genomes. Although Alphaherpesvirinae genomes are large and complicated to edit, in recent years several Pseudorabies virus (PRV) mutants have been successfully generated using the CRISPR\/Cas9 system. However, the application of CRISPR\/Cas9 editing on another member of alpha herpesviruses, bovine herpesvirus-1 (BHV-1), is rarely reported. This paper reports a rapid and straightforward approach to manipulating herpesviruses genome using CRISPR\/Cas9. The recombinant plasmids contained the left and right arm of the thymidine kinase (TK) gene of PRV or of the glycoprotein I (gI) and glycoprotein E (gE) of BHV-1. Upon the cleavage of the TK or gIgE gene by Cas9 protein, this was replaced by the enhanced green fluorescence protein (eGFP) by homologous recombination. With this approach, we generated recombinant TK-\/eGFP+ PRV and gIgE-\/eGFP+ BHV-1 mutants and then proceeded to characterize their biological activities in vitro and in vivo. In conclusion, we showed that alpha herpesvirus, including PRV and BHV-1, can be rapidly edited using the CRISPR\/Cas9 approach paving the way to the development of animal herpesvirus vaccines.<\/jats:p>","DOI":"10.3390\/v16020311","type":"journal-article","created":{"date-parts":[[2024,2,19]],"date-time":"2024-02-19T11:23:57Z","timestamp":1708341837000},"page":"311","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Application of CRISPR\/Cas9 for Rapid Genome Editing of Pseudorabies Virus and Bovine Herpesvirus-1"],"prefix":"10.3390","volume":"16","author":[{"given":"Wanqi","family":"Yu","sequence":"first","affiliation":[{"name":"Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, China"},{"name":"Institute of Animal Sciences, Wageningen University and Research, 6708 WD Wageningen, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jingyi","family":"Liu","sequence":"additional","affiliation":[{"name":"Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yingnan","family":"Liu","sequence":"additional","affiliation":[{"name":"Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9026-7320","authenticated-orcid":false,"given":"Maria","family":"Forlenza","sequence":"additional","affiliation":[{"name":"Institute of Animal Sciences, Wageningen University and Research, 6708 WD Wageningen, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7254-7596","authenticated-orcid":false,"given":"Hongjun","family":"Chen","sequence":"additional","affiliation":[{"name":"Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2024,2,18]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"429","DOI":"10.1016\/j.virol.2004.09.042","article-title":"Complete Genome Sequence of Cercopithecine Herpesvirus 2 (SA8) and Comparison with Other Simplexviruses","volume":"331","author":"Tyler","year":"2005","journal-title":"Virology"},{"key":"ref_2","unstructured":"Mahy, B.W.J., and Van Regenmortel, M.H.V. 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