Production of lentiviral vector with polycistronic transcripts for reprogramming of mouse fibroblast cells
iPS cells were originally generated using monocistronic retroviral vectors carrying the Yamanaka factors 'OSKM'. The development of a polycistronic viral vector with OSKM linked by 2A peptides has simplified reprogramming procedure and reduced the risk of multiple proviral integrations and...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Universiti Putra Malaysia Press
2018
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| Online Access: | http://psasir.upm.edu.my/id/eprint/66290/1/10%20JST%20Vol%2026%20%282%29%20Apr%202018_JST-0759-2016_pg627-640.pdf |
| _version_ | 1825933105612980224 |
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| author | Al Abbar, Akram Nordin, Norshariza Ngai, Siew Ching Abdullah, Syahrilnizam |
| author_facet | Al Abbar, Akram Nordin, Norshariza Ngai, Siew Ching Abdullah, Syahrilnizam |
| author_sort | Al Abbar, Akram |
| collection | UPM |
| description | iPS cells were originally generated using monocistronic retroviral vectors carrying the Yamanaka factors 'OSKM'. The development of a polycistronic viral vector with OSKM linked by 2A peptides has simplified reprogramming procedure and reduced the risk of multiple proviral integrations and insertional mutagenesis. In this study, we demonstrated the production of the polycistronic lentiviral vector encoding OSKM in a single cassette without a reporter gene or drug-based selection system. Syncytia formations were clearly seen following the co-transfection of a lentiviral plasmid construct with the structural and packaging plasmids. The virion was collected at 48 hours post-transfection. Afterwards, the viral titers were measured by the expression of Sox2 protein from transduced HT1080 cells. Subsequently, Oct4 expression was successfully detected in mouse fibroblasts in the range of 5, 10 and 20 MOIs with expression of 90.7%, 97.5% and 98%, respectively. The results obtained from this study could be used as a model for the production of OSKM lentiviral vector for newcomers to cellular reprogramming research. |
| first_indexed | 2024-03-06T09:52:11Z |
| format | Article |
| id | upm.eprints-66290 |
| institution | Universiti Putra Malaysia |
| language | English |
| last_indexed | 2024-03-06T09:52:11Z |
| publishDate | 2018 |
| publisher | Universiti Putra Malaysia Press |
| record_format | dspace |
| spelling | upm.eprints-662902019-02-12T07:01:57Z http://psasir.upm.edu.my/id/eprint/66290/ Production of lentiviral vector with polycistronic transcripts for reprogramming of mouse fibroblast cells Al Abbar, Akram Nordin, Norshariza Ngai, Siew Ching Abdullah, Syahrilnizam iPS cells were originally generated using monocistronic retroviral vectors carrying the Yamanaka factors 'OSKM'. The development of a polycistronic viral vector with OSKM linked by 2A peptides has simplified reprogramming procedure and reduced the risk of multiple proviral integrations and insertional mutagenesis. In this study, we demonstrated the production of the polycistronic lentiviral vector encoding OSKM in a single cassette without a reporter gene or drug-based selection system. Syncytia formations were clearly seen following the co-transfection of a lentiviral plasmid construct with the structural and packaging plasmids. The virion was collected at 48 hours post-transfection. Afterwards, the viral titers were measured by the expression of Sox2 protein from transduced HT1080 cells. Subsequently, Oct4 expression was successfully detected in mouse fibroblasts in the range of 5, 10 and 20 MOIs with expression of 90.7%, 97.5% and 98%, respectively. The results obtained from this study could be used as a model for the production of OSKM lentiviral vector for newcomers to cellular reprogramming research. Universiti Putra Malaysia Press 2018 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/66290/1/10%20JST%20Vol%2026%20%282%29%20Apr%202018_JST-0759-2016_pg627-640.pdf Al Abbar, Akram and Nordin, Norshariza and Ngai, Siew Ching and Abdullah, Syahrilnizam (2018) Production of lentiviral vector with polycistronic transcripts for reprogramming of mouse fibroblast cells. Pertanika Journal of Science & Technology, 26 (2). pp. 627-640. ISSN 0128-7680; ESSN: 2231-8526 http://www.pertanika.upm.edu.my/Pertanika%20PAPERS/JST%20Vol.%2026%20(2)%20Apr.%202018/10%20JST%20Vol%2026%20(2)%20Apr%202018_JST-0759-2016_pg627-640.pdf |
| spellingShingle | Al Abbar, Akram Nordin, Norshariza Ngai, Siew Ching Abdullah, Syahrilnizam Production of lentiviral vector with polycistronic transcripts for reprogramming of mouse fibroblast cells |
| title | Production of lentiviral vector with polycistronic transcripts for reprogramming of mouse fibroblast cells |
| title_full | Production of lentiviral vector with polycistronic transcripts for reprogramming of mouse fibroblast cells |
| title_fullStr | Production of lentiviral vector with polycistronic transcripts for reprogramming of mouse fibroblast cells |
| title_full_unstemmed | Production of lentiviral vector with polycistronic transcripts for reprogramming of mouse fibroblast cells |
| title_short | Production of lentiviral vector with polycistronic transcripts for reprogramming of mouse fibroblast cells |
| title_sort | production of lentiviral vector with polycistronic transcripts for reprogramming of mouse fibroblast cells |
| url | http://psasir.upm.edu.my/id/eprint/66290/1/10%20JST%20Vol%2026%20%282%29%20Apr%202018_JST-0759-2016_pg627-640.pdf |
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