A transient three-plasmid expression system for the production of high titer retroviral vectors (original) (raw)

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Retrovirus Molecular Biology Group, Department of Biochemistry, University of Oxford

South Parks Road, Oxford 0X1 3QU, UK

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Retrovirus Molecular Biology Group, Department of Biochemistry, University of Oxford

South Parks Road, Oxford 0X1 3QU, UK

Search for other works by this author on:

,

Retrovirus Molecular Biology Group, Department of Biochemistry, University of Oxford

South Parks Road, Oxford 0X1 3QU, UK

Search for other works by this author on:

,

Retrovirus Molecular Biology Group, Department of Biochemistry, University of Oxford

South Parks Road, Oxford 0X1 3QU, UK

Search for other works by this author on:

,

Retrovirus Molecular Biology Group, Department of Biochemistry, University of Oxford

South Parks Road, Oxford 0X1 3QU, UK

Search for other works by this author on:

,

Retrovirus Molecular Biology Group, Department of Biochemistry, University of Oxford

South Parks Road, Oxford 0X1 3QU, UK

Search for other works by this author on:

Retrovirus Molecular Biology Group, Department of Biochemistry, University of Oxford

South Parks Road, Oxford 0X1 3QU, UK

* To whom correspondence should be addressed

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Received:

19 October 1994

Revision received:

11 January 1995

Accepted:

11 January 1995

Published:

25 February 1995

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Yuko Soneoka, Paula M. Cannon, Emma E. Ramsdale, Joanne C. Griffiths, Gaetano Romano, Susan M. Kingsman, Alan J. Kingsman, A transient three-plasmid expression system for the production of high titer retroviral vectors, Nucleic Acids Research, Volume 23, Issue 4, 25 February 1995, Pages 628–633, https://doi.org/10.1093/nar/23.4.628
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Abstract

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 ∼10 7 Infectious units/ml by 48 h after transient co-transfectlon with the three plasmid components. This system can be used both for the generation of high titer retroviral stocks for transductlon and for the rapid screening of a large number of MLV gag-pol or env mutants.

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