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pCOR: a new design of plasmid vectors for nonviral gene therapy

Gene Therapy volume 6, pages 1482–1488 (1999)Cite this article

Abstract

A totally redesigned host/vector system with improved properties in terms of safety has been developed. The pCOR plasmids are narrow-host range plasmid vectors for nonviral gene therapy. These plasmids contain a conditional origin of replication and must be propagated in a specifically engineered E. coli host strain, greatly reducing the potential for propagation in the environment or in treated patients. The pCOR backbone has several features that increase safety in terms of dissemination and selection: (1) the origin of replication requires a plasmid-specific initiator protein, π protein, encoded by the pir gene limiting its host range to bacterial strains that produce this trans-acting protein; (2) the plasmid’s selectable marker is not an antibiotic resistance gene but a gene encoding a bacterial suppressor tRNA. Optimized E. coli hosts supporting pCOR replication and selection were constructed. High yields of supercoiled pCOR monomers were obtained (100 mg/l) through fed-batch fermentation. pCOR vectors carrying the luciferase reporter gene gave high levels of luciferase activity when injected into murine skeletal muscle.

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Acknowledgements

We would like especially to thank JM Masson for providing sup Phe and XAC-1 and B Wanner for ori γ and uidA: :pir cassettes. W Wackernagel is acknowledged for the generous gift of the endA fragment. We would like to thank F Blanche and collaborators for topoisomerase experiments and T Ciora for oligonucleotide synthesis and sequencing.

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  1. Rhône-Poulenc Rorer, Centre de Recherche de Vitry Alfortville, 13 Quai J Guesde, Vitry-sur-Seine, 94403, France

    F Soubrier, B Cameron, B Manse, S Somarriba, C Dubertret, G Jaslin, G Jung, C Le Caer, D Dang, J M Mouvault, D Scherman, J F Mayaux & J Crouzet

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  1. F Soubrier
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Soubrier, F., Cameron, B., Manse, B. et al. pCOR: a new design of plasmid vectors for nonviral gene therapy. Gene Ther 6, 1482–1488 (1999). https://doi.org/10.1038/sj.gt.3300968

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