RSAD2 Is an Effective Target for High-Yield Vaccine Production in MDCK Cells
Increasingly, attention has focused on improving vaccine production in cells using gene editing technology to specifically modify key virus regulation-related genes to promote virus replication. In this study, we used DIA proteomics analysis technology to compare protein expression differences betwe...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-11-01
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| Series: | Viruses |
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| Online Access: | https://www.mdpi.com/1999-4915/14/11/2587 |
| _version_ | 1797463801287671808 |
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| author | Zilin Qiao Yuejiao Liao Mengyuan Pei Zhenyu Qiu Zhenbin Liu Dongwu Jin Jiayou Zhang Zhongren Ma Xiaoming Yang |
| author_facet | Zilin Qiao Yuejiao Liao Mengyuan Pei Zhenyu Qiu Zhenbin Liu Dongwu Jin Jiayou Zhang Zhongren Ma Xiaoming Yang |
| author_sort | Zilin Qiao |
| collection | DOAJ |
| description | Increasingly, attention has focused on improving vaccine production in cells using gene editing technology to specifically modify key virus regulation-related genes to promote virus replication. In this study, we used DIA proteomics analysis technology to compare protein expression differences between two groups of MDCK cells: uninfected and influenza A virus (IAV) H1N1-infected cells 16 h post infection (MOI = 0.01). Initially, 266 differentially expressed proteins were detected after infection, 157 of which were upregulated and 109 were downregulated. We screened these proteins to 23 genes related to antiviral innate immunity regulation based on functional annotation database analysis and verified the mRNA expression of these genes using qPCR. Combining our results with published literature, we focused on the proteins RSAD2, KCNN4, IDO1, and ISG20; we verified their expression using western blot, which was consistent with our proteomics results. Finally, we knocked down RSAD2 using lentiviral shRNA expression vectors and found that RSAD2 inhibition significantly increased IAV NP gene expression, effectively promoting influenza virus replication with no significant effect on cell proliferation. These results indicate that RSAD2 is potentially an effective target for establishing high-yield vaccine MDCK cell lines and will help to fully understand the interaction mechanism between host cells and influenza viruses. |
| first_indexed | 2024-03-09T17:55:53Z |
| format | Article |
| id | doaj.art-1702d9ce028e48ec95ca4d603b2b1b03 |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-09T17:55:53Z |
| publishDate | 2022-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-1702d9ce028e48ec95ca4d603b2b1b032023-11-24T10:19:39ZengMDPI AGViruses1999-49152022-11-011411258710.3390/v14112587RSAD2 Is an Effective Target for High-Yield Vaccine Production in MDCK CellsZilin Qiao0Yuejiao Liao1Mengyuan Pei2Zhenyu Qiu3Zhenbin Liu4Dongwu Jin5Jiayou Zhang6Zhongren Ma7Xiaoming Yang8Gansu Tech Innovation Center of Animal Cell, Biomedical Research Center, Northwest Minzu University, Lanzhou 730030, ChinaGansu Tech Innovation Center of Animal Cell, Biomedical Research Center, Northwest Minzu University, Lanzhou 730030, ChinaGansu Tech Innovation Center of Animal Cell, Biomedical Research Center, Northwest Minzu University, Lanzhou 730030, ChinaGansu Tech Innovation Center of Animal Cell, Biomedical Research Center, Northwest Minzu University, Lanzhou 730030, ChinaGansu Tech Innovation Center of Animal Cell, Biomedical Research Center, Northwest Minzu University, Lanzhou 730030, ChinaGansu Provincial Bioengineering Materials Engineering Research Center, Lanzhou Minhai Bio-Engineering Co., Ltd., Lanzhou 730030, ChinaNational Engineering Technology Research Center for Combined Vaccines, Wuhan 430207, ChinaGansu Tech Innovation Center of Animal Cell, Biomedical Research Center, Northwest Minzu University, Lanzhou 730030, ChinaNational Engineering Technology Research Center for Combined Vaccines, Wuhan 430207, ChinaIncreasingly, attention has focused on improving vaccine production in cells using gene editing technology to specifically modify key virus regulation-related genes to promote virus replication. In this study, we used DIA proteomics analysis technology to compare protein expression differences between two groups of MDCK cells: uninfected and influenza A virus (IAV) H1N1-infected cells 16 h post infection (MOI = 0.01). Initially, 266 differentially expressed proteins were detected after infection, 157 of which were upregulated and 109 were downregulated. We screened these proteins to 23 genes related to antiviral innate immunity regulation based on functional annotation database analysis and verified the mRNA expression of these genes using qPCR. Combining our results with published literature, we focused on the proteins RSAD2, KCNN4, IDO1, and ISG20; we verified their expression using western blot, which was consistent with our proteomics results. Finally, we knocked down RSAD2 using lentiviral shRNA expression vectors and found that RSAD2 inhibition significantly increased IAV NP gene expression, effectively promoting influenza virus replication with no significant effect on cell proliferation. These results indicate that RSAD2 is potentially an effective target for establishing high-yield vaccine MDCK cell lines and will help to fully understand the interaction mechanism between host cells and influenza viruses.https://www.mdpi.com/1999-4915/14/11/2587DIAMDCKinfluenza virusRSAD2vaccine |
| spellingShingle | Zilin Qiao Yuejiao Liao Mengyuan Pei Zhenyu Qiu Zhenbin Liu Dongwu Jin Jiayou Zhang Zhongren Ma Xiaoming Yang RSAD2 Is an Effective Target for High-Yield Vaccine Production in MDCK Cells Viruses DIA MDCK influenza virus RSAD2 vaccine |
| title | RSAD2 Is an Effective Target for High-Yield Vaccine Production in MDCK Cells |
| title_full | RSAD2 Is an Effective Target for High-Yield Vaccine Production in MDCK Cells |
| title_fullStr | RSAD2 Is an Effective Target for High-Yield Vaccine Production in MDCK Cells |
| title_full_unstemmed | RSAD2 Is an Effective Target for High-Yield Vaccine Production in MDCK Cells |
| title_short | RSAD2 Is an Effective Target for High-Yield Vaccine Production in MDCK Cells |
| title_sort | rsad2 is an effective target for high yield vaccine production in mdck cells |
| topic | DIA MDCK influenza virus RSAD2 vaccine |
| url | https://www.mdpi.com/1999-4915/14/11/2587 |
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