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ORIGINAL RESEARCH article
Front. Microbiol.
Sec. Virology
Volume 15 - 2024 |
doi: 10.3389/fmicb.2024.1483527
IRF7-deficient MDBK cell based on CRISPR/Cas9 technology for enhancing IBRV replication
Provisionally accepted- 1 College of Animal Science and Technology, Jilin Agricultural University, Changchun, China
- 2 Wengniute banner Agriculture and Animal Husbandry Bureau, Chifeng, China
- 3 College of Chinese Medicine Materials, Jilin Agricultural University, Changchun, China
Infectious bovine rhinotracheitis (IBR), characterized by acute respiratory lesions in cattle, is a major infectious disease caused by bovine alphaherpesvirus-1 (BoAHV-1). Control of this disease is primarily depending on vaccination. Madin-Darby bovine kidney cells (MDBK) being the main host cells and the important production platform for IBR vaccines. However, innate immune genes inhibit viral replication. Accordingly, the aim of this study was developing of IRF7 gene deleted MDBK cells to facilitate the production of high-titer vaccines. The CRISPR/Cas9 technology was used to knock out the IRF7 gene in MDBK cells and the impact on virus replication was examined using virus growth curves, CCK-8 assays, cell scratch assays, and qPCR. The knockout of the IRF7 gene in MDBK cells led to an increased replication capacity of IBRV and a significant reduction in type I interferons expression, specifically IFN-α and IFN-β. This indicates that IRF7 -/-MDBK cell lines can effectively result in production of IBRV with high-titer, which will enhance the development of inactivated or attenuated vaccines.
Keywords: CRISPR/Cas9, gene knockout, IRF7, IBRV, viral replication
Received: 20 Aug 2024; Accepted: 05 Nov 2024.
Copyright: © 2024 Ge, Dongli, Ling, Xu, Wang, Li, Hao, Gong, Li, Shi, Leng and Du. 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:
Qian Ling, College of Chinese Medicine Materials, Jilin Agricultural University, Changchun, China
Hui Li Xu, College of Animal Science and Technology, Jilin Agricultural University, Changchun, China
Lin Xiao Wang, College of Animal Science and Technology, Jilin Agricultural University, Changchun, China
Yan Ke Li, College of Animal Science and Technology, Jilin Agricultural University, Changchun, China
Wen Hao, College of Animal Science and Technology, Jilin Agricultural University, Changchun, China
Xue Leng, College of Chinese Medicine Materials, Jilin Agricultural University, Changchun, China
Rui Du, College of Chinese Medicine Materials, Jilin Agricultural University, Changchun, China
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