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A transient three-plasmid expression system for the production of high titer retroviral vectors
Open accessAbstract
We have constructed a series of MLV-based retroviral vectors and packaging components expressed from the CMV promoter and carried on plasmids containing SV40 origins of replication. These two features greatly enhanced retroviral gene expression when introduced into cell lines carrying the SV40 large T antigen. The two packaging components, gag-pol and env, were placed on separate plasmids to reduce helper virus formation. Using a highly transfectable human cell line and sodium butyrate to further increase expression of each component, we achieved helper-free viral stocks of approximately 10(7) infectious units/ml by 48 h after transient co-transfection with the three plasmid components. This system can be used both for the generation of high titer retroviral stocks for transduction and for the rapid screening of a large number of MLV gag-pol or env mutants.
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Cited by
In Vivo Gene Delivery and Stable Transduction of Nondividing Cells by a Lentiviral Vector
Naldini, L., Blömer, U., Gallay, P., Ory, D., Mulligan, R., Gage, F. H., Verma, I. M., & Trono, D. (1996) · Science · doi:10.1126/science.272.5259.263 · Cited by 4,903
Goldmacher, V. S., Bartle, L. M., Skaletskaya, A., Dionne, C. A., Kedersha, N. L., Vater, C. A., Han, J.-W., Lutz, R. J., Watanabe, S., McFarland, E. D. C., et al. (1999) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.96.22.12536 · Cited by 423
Duensing, S., Lee, L. Y., Duensing, A., Basile, J., Piboonniyom, S.-O., Gonzalez, S., Crum, C. P., & Münger, K. (2000) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.170093297 · Cited by 397
Fouchier, R. A. M. (1997) · The EMBO Journal · doi:10.1093/emboj/16.15.4531 · Cited by 339
Srivastava, S., Salter, A. I., Liggitt, D., Yechan-Gunja, S., Sarvothama, M., Cooper, K., Smythe, K. S., Dudakov, J. A., Pierce, R. H., Rader, C., et al. (2019) · Cancer Cell · doi:10.1016/j.ccell.2019.02.003 · Cited by 334
Metadata
- Authors
- Soneoka, Y., Cannon, P. M., Ramsdale, E. E., Griffiths, J. C., Romano, G., Kingsman, S. M., & Kingsman, A. J.
- Venue
- Nucleic Acids Research
- Publisher
- Oxford University Press (OUP)
- Published
- 1995
- Volume
- 23
- Issue
- 4
- Pages
- 628-633
- ISSN
- 0305-1048 (ISSN-L)
- 1362-4962
- OpenAlex
- W2147153603
- Cited by
- 690
- References
- 0
- FWCI
- 9.94
- Type
- journal article
- Language
- en
- Open access
- green
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- Soneoka, Y., Cannon, P. M., Ramsdale, E. E., Griffiths, J. C., Romano, G., Kingsman, S. M., & Kingsman, A. J. (1995). A transient three-plasmid expression system for the production of high titer retroviral vectors. Nucleic Acids Research. https://doi.org/10.1093/nar/23.4.628
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