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Insulators coupled to a minimal bidirectional tet cassette for tight regulation of rAAV-mediated gene transfer in the mammalian brain

Gene Therapy volume 8, pages 1675–1681 (2001)Cite this article

Abstract

Recombinant AAV is increasingly becoming the vector of choice for many gene therapy applications in the CNS, due to its lack of toxicity and high level of sustained expression. With recent improvements in the generation of pure, high titer vector stocks, the regulation of gene expression is now a key issue for successful translation of gene therapy-based treatments to the clinic. The level of the transgene protein may need to be maintained within a narrow therapeutic window for the successful treatment of human disease. The doxycycline responsive system directs a dose-responsive, tightly regulated level of gene expression and has been used successfully in transgenic mouse models. Here, we have optimized an autoregulatory, bidirectional doxycyline responsive cassette specifically for use in rAAV. We minimized the size of the cassette and decreased the basal leakiness of the system, leading to tight regulation in the rat brain.

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Acknowledgements

We thank R Bland, D Young and T Lawlor for critical reading of the manuscript. We are grateful to C Strathdee for providing the pBIG3r plasmid. This work was supported by the New Economy Research Fund, New Zealand Health Research Council, the National Institutes of Health and the Jefferson Faculty Foundation.

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Authors and Affiliations

  1. Division of Molecular Medicine, Faculty of Medical and Health Sciences, University of Auckland, Auckland, New Zealand

    HL Fitzsimons, JM McKenzie & MJ During

  2. CNS Gene Therapy Center, Jefferson Medical College, Philadelphia, PA, USA

    MJ During

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  1. HL Fitzsimons
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  2. JM McKenzie
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  3. MJ During
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Fitzsimons, H., McKenzie, J. & During, M. Insulators coupled to a minimal bidirectional tet cassette for tight regulation of rAAV-mediated gene transfer in the mammalian brain. Gene Ther 8, 1675–1681 (2001). https://doi.org/10.1038/sj.gt.3301582

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