Technical Report | Published:

Chemical control of protein stability and function in living mice

Nature Medicine volume 14, pages 11231127 (2008) | Download Citation

Abstract

Conditional control of protein function in vivo offers great potential for deconvoluting the roles of individual proteins in complicated systems. We recently developed a method in which a small protein domain, termed a destabilizing domain, confers instability to fusion protein partners in cultured cells. Instability is reversed when a cell-permeable small molecule binds this domain. Here we describe the use of this system to regulate protein function in living mammals. We show regulation of secreted proteins and their biological activity with conditional secretion of an immunomodulatory cytokine, resulting in tumor burden reduction in mouse models. Additionally, we use this approach to control the function of a specific protein after systemic delivery of the gene that encodes it to a tumor, suggesting uses for enhancing the specificity and efficacy of targeted gene-based therapies. This method represents a new strategy to regulate protein function in living organisms with a high level of control.

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Acknowledgements

This work was supported by the US National Institutes of Health, through the Small Animal Imaging Resource Program, the In vivo Cellular and Molecular Imaging Center (grant numbers R24 CA92862 and P50 CA114747) and GM073046, and the John A. and Cynthia Fry Gunn Research Fund. We thank T. Clackson for the Shield-1 formulation protocol and B. Moss (US National Institutes of Health) for pSC-65 and VSC20.

Author information

Author notes

    • Steve H Thorne

    Current address: University of Pittsburgh Cancer Institute and Department of Surgery, University of Pittsburgh, 5117 Centre Avenue, Pittsburgh, Pennsylvania 15213, USA.

    • Laura A Banaszynski
    •  & Mark A Sellmyer

    These authors contributed equally to this work.

Affiliations

  1. Department of Chemistry, Radiology and Microbiology & Immunology, Stanford University, 318 Campus Drive, Stanford, California 94305, USA.

    • Laura A Banaszynski
  2. Department of Chemical & Systems Biology, Radiology and Microbiology & Immunology, Stanford University, 318 Campus Drive, Stanford, California 94305, USA.

    • Mark A Sellmyer
    •  & Thomas J Wandless
  3. Departments of Pediatrics, Radiology and Microbiology & Immunology, Stanford University, 318 Campus Drive, Stanford, California 94305, USA.

    • Mark A Sellmyer
    • , Christopher H Contag
    •  & Steve H Thorne

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Contributions

L.A.B. and M.A.S. designed and performed research and analyzed data. S.H.T. designed, coordinated and performed research and analyzed data. C.H.C. and T.J.W. helped design and interpret experiments and analyze data. All authors wrote the manuscript.

Competing interests

C.H.C. is a founder of Xenogen—now Caliper Life Sciences—and a consultant for Caliper Life Sciences.

Corresponding authors

Correspondence to Christopher H Contag or Thomas J Wandless.

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DOI

https://doi.org/10.1038/nm.1754

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