BRIEF RESEARCH REPORT article
Front. Cell. Infect. Microbiol.
Sec. Veterinary and Zoonotic Infection
Volume 15 - 2025 | doi: 10.3389/fcimb.2025.1660446
This article is part of the Research TopicUnveiling Host-Pathogen Interactions: Insights into Animal Cellular Immunity and Novel Diagnostics - Volume IIView all 18 articles
Highly Efficient Gene Editing of Feline Herpesvirus 1 (FHV-1) Using CRISPR/Cas9 Combined with FACS
Provisionally accepted
- 1Jiangsu Agri-animal Husbandry Vocational College, Taizhou, China
- 2Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, China
- 3College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, China
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Feline herpesvirus 1 (FHV-1) is a major causative agent of feline viral rhinotracheitis and ocular lesions. Due to its large DNA genome, the construction of recombinant FHV-1 viruses presents considerable challenges for conventional methodologies. In this study, we implemented an integrated strategy combining CRISPR/Cas9-mediated gene editing with fluorescence-activated cell sorting (FACS) to enable the rapid and efficient generation of recombinant FHV-1 viruses. Specifically, the thymidine kinase (tk) gene was disrupted by inserting a monomeric Cherry (mCherry) reporter gene, and the glycoprotein E (gE) gene was similarly interrupted through the incorporation of a green fluorescent protein (GFP) reporter. The CRISPR/Cas9 system enables precise, sitespecific genomic modifications, while FACS allows for effective enrichment and isolation of the desired recombinant viral populations. This combined approach significantly reduces the time required for recombinant virus generation from weeks to days, thereby offering substantial potential to expedite vaccine development and advance functional genomics research.
Keywords: Feline herpesvirus 1 (FHV-1), CRISPR/Cas9, Fluorescence-activated cell sorting (FACS), Homologous Recombination, gene knockout
Received: 06 Jul 2025; Accepted: 06 Aug 2025.
Copyright: © 2025 Wang, Xu, Cai, Qiu, Lu and Tang. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Yan-Dong Tang, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, China
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