Generation of a bovine cell line for gene engineering using an HIV-1-based lentiviral vector
Abstract Human immunodeficiency virus type 1 (HIV-1)-based lentiviral vectors are indispensable tools for gene engineering in mammalian cells. Conversely, lentiviral vector transduction is severely inhibited in bovine cells. Previous studies demonstrated that this inhibition is caused by the anti-le...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2022-10-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-022-20970-6 |
| _version_ | 1797986066343395328 |
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| author | Nanami Morizako Erika P. Butlertanaka Yuri L. Tanaka Honoka Shibata Tamaki Okabayashi Hirohisa Mekata Akatsuki Saito |
| author_facet | Nanami Morizako Erika P. Butlertanaka Yuri L. Tanaka Honoka Shibata Tamaki Okabayashi Hirohisa Mekata Akatsuki Saito |
| author_sort | Nanami Morizako |
| collection | DOAJ |
| description | Abstract Human immunodeficiency virus type 1 (HIV-1)-based lentiviral vectors are indispensable tools for gene engineering in mammalian cells. Conversely, lentiviral vector transduction is severely inhibited in bovine cells. Previous studies demonstrated that this inhibition is caused by the anti-lentiviral host factor tripartite motif containing 5 (TRIM5), which targets incoming HIV-1 virions by interacting with the viral capsid. In this study, we investigated several methods for overcoming the limited applicability of lentiviral vectors in bovine cells. First, we demonstrated that the SPRY domain of bovine TRIM5 is the major determinant of anti-viral activity. Second, we found that mutations that allow the capsid to evade rhesus macaque TRIM5α minimally rescued HIV-1 infectivity in bovine-derived MDBK cells. Third, we found that cyclosporine A, which relieves the inhibition of HIV-1 infection in monkey cells, significantly rescued the impaired HIV-1 infectivity in MDBK cells. Lastly, we successfully generated a bovine cell line lacking intact TRIM5 using the CRISPR/Cas9 technique. This TRIM5 knockout cell line displayed significantly higher susceptibility to an HIV-1-based lentiviral vector. In conclusion, our findings provide a promising gene engineering strategy for bovine cells, thereby contributing to innovations in agriculture and improvements in animal health. |
| first_indexed | 2024-04-11T07:28:11Z |
| format | Article |
| id | doaj.art-fb5a28d69407433d9554ccf73471a100 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-04-11T07:28:11Z |
| publishDate | 2022-10-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-fb5a28d69407433d9554ccf73471a1002022-12-22T04:37:01ZengNature PortfolioScientific Reports2045-23222022-10-0112111410.1038/s41598-022-20970-6Generation of a bovine cell line for gene engineering using an HIV-1-based lentiviral vectorNanami Morizako0Erika P. Butlertanaka1Yuri L. Tanaka2Honoka Shibata3Tamaki Okabayashi4Hirohisa Mekata5Akatsuki Saito6Department of Veterinary Science, Faculty of Agriculture, University of MiyazakiDepartment of Veterinary Science, Faculty of Agriculture, University of MiyazakiDepartment of Veterinary Science, Faculty of Agriculture, University of MiyazakiDepartment of Veterinary Science, Faculty of Agriculture, University of MiyazakiDepartment of Veterinary Science, Faculty of Agriculture, University of MiyazakiCenter for Animal Disease Control, University of MiyazakiDepartment of Veterinary Science, Faculty of Agriculture, University of MiyazakiAbstract Human immunodeficiency virus type 1 (HIV-1)-based lentiviral vectors are indispensable tools for gene engineering in mammalian cells. Conversely, lentiviral vector transduction is severely inhibited in bovine cells. Previous studies demonstrated that this inhibition is caused by the anti-lentiviral host factor tripartite motif containing 5 (TRIM5), which targets incoming HIV-1 virions by interacting with the viral capsid. In this study, we investigated several methods for overcoming the limited applicability of lentiviral vectors in bovine cells. First, we demonstrated that the SPRY domain of bovine TRIM5 is the major determinant of anti-viral activity. Second, we found that mutations that allow the capsid to evade rhesus macaque TRIM5α minimally rescued HIV-1 infectivity in bovine-derived MDBK cells. Third, we found that cyclosporine A, which relieves the inhibition of HIV-1 infection in monkey cells, significantly rescued the impaired HIV-1 infectivity in MDBK cells. Lastly, we successfully generated a bovine cell line lacking intact TRIM5 using the CRISPR/Cas9 technique. This TRIM5 knockout cell line displayed significantly higher susceptibility to an HIV-1-based lentiviral vector. In conclusion, our findings provide a promising gene engineering strategy for bovine cells, thereby contributing to innovations in agriculture and improvements in animal health.https://doi.org/10.1038/s41598-022-20970-6 |
| spellingShingle | Nanami Morizako Erika P. Butlertanaka Yuri L. Tanaka Honoka Shibata Tamaki Okabayashi Hirohisa Mekata Akatsuki Saito Generation of a bovine cell line for gene engineering using an HIV-1-based lentiviral vector Scientific Reports |
| title | Generation of a bovine cell line for gene engineering using an HIV-1-based lentiviral vector |
| title_full | Generation of a bovine cell line for gene engineering using an HIV-1-based lentiviral vector |
| title_fullStr | Generation of a bovine cell line for gene engineering using an HIV-1-based lentiviral vector |
| title_full_unstemmed | Generation of a bovine cell line for gene engineering using an HIV-1-based lentiviral vector |
| title_short | Generation of a bovine cell line for gene engineering using an HIV-1-based lentiviral vector |
| title_sort | generation of a bovine cell line for gene engineering using an hiv 1 based lentiviral vector |
| url | https://doi.org/10.1038/s41598-022-20970-6 |
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