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| Effect of Vesicle Transport Protein SEC22B Knockdown on the Replication of Bovine viral diarrhea virus |
| ZHANG Cheng-Yuan, YUAN Yuan-Yuan, GUO Yan-Ting, YANG Li, RAN Duo-Liang, FU Qiang, SHI Hui-Jun* |
| College of Animal Medicine, Xinjiang Agricultural University, Urumqi 830052, China |
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Abstract The infection of bovine viral diarrhea is becoming more and more serious, which has caused serious losses to the breeding industry. At present, the lack of effective treatment leads to the complicated infection pattern in China, and the research on the pathogenesis and virus-host interaction is still unclear. To investigate the effect of vesicle transport protein (SEC22B) deletion on Bovine viral diarrhea virus (BVDV) replication, according to the SEC22B gene sequence in GenBank database, small guide RNA (sgRNA) was designed using Benchling platform and cloned into LentiCRISPR V2 vector for lentivirus packaging. The SEC22B gene in madin-darby bovine kidney cells (MDBK) was knocked down (KD) by CRISPR/Cas9 gene editing technique, and identified by Western blot. After BVDV infection with SEC22B KD cells at different times, the changes of 5' UTR mRNA level and accumulation of double-stranded RNA (dsRNA) were detected by qPCR and immunofluorescence staining, respectively. Reed-Muench method was used to measure the titer change of progeny virus after BVDV infection at different time. The results showed that compared with scramble cells, the protein expression level of SEC22B in SEC22B KD cells was significantly decreased by Western blot. Compared with scramble cells infected with BVDV, the level of 5' UTR mRNA and dsRNA of SEC22B KD cells were extremely significantly decreased (P<0.01) after BVDV infection, and the titer of progeny virus was extremely significantly decreased (P<0.01). The results showed that knockdown SEC22B gene significantly inhibited the replication of BVDV. This study provides a target for the establishment of new methods for the prevention and control of BVDV.
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Received: 14 April 2022
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Corresponding Authors:
* shihuijunmm@163.com
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