Development of a T7 RNA polymerase expressing cell line using lentivirus vectors for the recovery of recombinant Newcastle disease virus
The development of a T7 RNA polymerase (T7 RNAP) expressing cell line i.e. BSR T7/5 cells marks an improvement of reverse genetics for the recovery of recombinant Newcastle disease virus (rNDV). BSR T7/5 is developed by transient transfection of plasmid encoding T7 RNAP gene for rNDV rescue. However...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2021
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| Online Access: | http://psasir.upm.edu.my/id/eprint/96691/1/ABSTRACT.pdf |
| _version_ | 1825937750032908288 |
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| author | Yeong, Ming Yue Cheow, Pheik Sheen Abdullah, Syahril Song, Adelene Ai Lian Lei, Janet Rossmann Tan, Tiong Kit Yusoff, Khatijah Chia, Suet Lin |
| author_facet | Yeong, Ming Yue Cheow, Pheik Sheen Abdullah, Syahril Song, Adelene Ai Lian Lei, Janet Rossmann Tan, Tiong Kit Yusoff, Khatijah Chia, Suet Lin |
| author_sort | Yeong, Ming Yue |
| collection | UPM |
| description | The development of a T7 RNA polymerase (T7 RNAP) expressing cell line i.e. BSR T7/5 cells marks an improvement of reverse genetics for the recovery of recombinant Newcastle disease virus (rNDV). BSR T7/5 is developed by transient transfection of plasmid encoding T7 RNAP gene for rNDV rescue. However, the gene expression decreases gradually over multiple passages and eventually hinders the rescue of rNDV. To address this issue, lentiviral vector was used to develop T7 RNAP-expressing HEK293-TA (HEK293-TA-Lv-T7) and SW620 (SW620-Lv-T7) cell lines, evidenced by the expression of T7 RNAP after subsequent 20 passages. rNDV was rescued successfully using HEK293-TA-Lv-T7 clones (R1D3, R1D8, R5B9) and SW620-Lv-T7 clones (R1C11, R3C5) by reverse transfection, yielding comparable virus rescue efficiency and virus titres to that of BSR T7/5. This study provides new tools for rNDV rescue and insights into cell line development and virology by reverse genetics. |
| first_indexed | 2024-03-06T11:04:16Z |
| format | Article |
| id | upm.eprints-96691 |
| institution | Universiti Putra Malaysia |
| language | English |
| last_indexed | 2024-03-06T11:04:16Z |
| publishDate | 2021 |
| publisher | Elsevier |
| record_format | dspace |
| spelling | upm.eprints-966912022-12-14T13:16:46Z http://psasir.upm.edu.my/id/eprint/96691/ Development of a T7 RNA polymerase expressing cell line using lentivirus vectors for the recovery of recombinant Newcastle disease virus Yeong, Ming Yue Cheow, Pheik Sheen Abdullah, Syahril Song, Adelene Ai Lian Lei, Janet Rossmann Tan, Tiong Kit Yusoff, Khatijah Chia, Suet Lin The development of a T7 RNA polymerase (T7 RNAP) expressing cell line i.e. BSR T7/5 cells marks an improvement of reverse genetics for the recovery of recombinant Newcastle disease virus (rNDV). BSR T7/5 is developed by transient transfection of plasmid encoding T7 RNAP gene for rNDV rescue. However, the gene expression decreases gradually over multiple passages and eventually hinders the rescue of rNDV. To address this issue, lentiviral vector was used to develop T7 RNAP-expressing HEK293-TA (HEK293-TA-Lv-T7) and SW620 (SW620-Lv-T7) cell lines, evidenced by the expression of T7 RNAP after subsequent 20 passages. rNDV was rescued successfully using HEK293-TA-Lv-T7 clones (R1D3, R1D8, R5B9) and SW620-Lv-T7 clones (R1C11, R3C5) by reverse transfection, yielding comparable virus rescue efficiency and virus titres to that of BSR T7/5. This study provides new tools for rNDV rescue and insights into cell line development and virology by reverse genetics. Elsevier 2021 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/96691/1/ABSTRACT.pdf Yeong, Ming Yue and Cheow, Pheik Sheen and Abdullah, Syahril and Song, Adelene Ai Lian and Lei, Janet Rossmann and Tan, Tiong Kit and Yusoff, Khatijah and Chia, Suet Lin (2021) Development of a T7 RNA polymerase expressing cell line using lentivirus vectors for the recovery of recombinant Newcastle disease virus. Journal of Virological Methods, 291. art. no. 114099. pp. 1-9. ISSN 0166-0934; ESSN: 879-0984 https://www.sciencedirect.com/science/article/pii/S0166093421000380 10.1016/j.jviromet.2021.114099 |
| spellingShingle | Yeong, Ming Yue Cheow, Pheik Sheen Abdullah, Syahril Song, Adelene Ai Lian Lei, Janet Rossmann Tan, Tiong Kit Yusoff, Khatijah Chia, Suet Lin Development of a T7 RNA polymerase expressing cell line using lentivirus vectors for the recovery of recombinant Newcastle disease virus |
| title | Development of a T7 RNA polymerase expressing cell line using lentivirus vectors for the recovery of recombinant Newcastle disease virus |
| title_full | Development of a T7 RNA polymerase expressing cell line using lentivirus vectors for the recovery of recombinant Newcastle disease virus |
| title_fullStr | Development of a T7 RNA polymerase expressing cell line using lentivirus vectors for the recovery of recombinant Newcastle disease virus |
| title_full_unstemmed | Development of a T7 RNA polymerase expressing cell line using lentivirus vectors for the recovery of recombinant Newcastle disease virus |
| title_short | Development of a T7 RNA polymerase expressing cell line using lentivirus vectors for the recovery of recombinant Newcastle disease virus |
| title_sort | development of a t7 rna polymerase expressing cell line using lentivirus vectors for the recovery of recombinant newcastle disease virus |
| url | http://psasir.upm.edu.my/id/eprint/96691/1/ABSTRACT.pdf |
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