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Drug-inducible and simultaneous regulation of endogenous genes by single-chain nuclear receptor-based zinc-finger transcription factor gene switches

Gene Therapy volume 15, pages 1223–1232 (2008)Cite this article

An Erratum to this article was published on 19 August 2008

Abstract

Chemically inducible gene switches that regulate expression of endogenous genes have multiple applications for basic gene expression research and gene therapy. Single-chain zinc-finger transcription factors that utilize either estrogen receptor homodimers or retinoid X receptor-α/ecdysone receptor heterodimers are shown here to be effective regulators of ICAM-1 and ErbB-2 transcription. Using activator (VP64) and repressor (Krüppel-associated box) domains to impart regulatory directionality, ICAM-1 was activated by 4.8-fold and repressed by 81% with the estrogen receptor-inducible transcription factors. ErbB-2 was activated by up to threefold and repressed by 84% with the retinoid X receptor-α/ecdysone receptor-inducible transcription factors. The dynamic range of these proteins was similar to the constitutive system and showed negligible basal regulation when ligand was not present. We have also demonstrated that the regulation imposed by these inducible transcription factors is dose dependent, sustainable for at least 11 days and reversible upon cessation of drug treatment. Importantly, these proteins can be used in conjunction with each other with no detectable overlap of activity enabling concurrent and temporal regulation of multiple genes within the same cell. Thus, these chemically inducible transcription factors are valuable tools for spatiotemporal control of gene expression that should prove valuable for research and gene therapy applications.

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Acknowledgements

This study was supported in part by the National Institute of Health Grant GM065059 to CFB. LM was the recipient of postdoctoral fellowships from the Swiss National Science Foundation. LJS is supported by The American Cancer Society Illinois Division—Linda M Campbell Postdoctoral Fellowship.

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  1. L Magnenat

    Present address: 4Current address: Merck Serono Geneva Research Center, Geneva, Switzerland.,

Authors and Affiliations

  1. The Skaggs Institute for Chemical Biology, La Jolla, CA, USA

    L Magnenat, L J Schwimmer & C F Barbas III

  2. Department of Molecular Biology, The Scripps Research Institute, La Jolla, CA, USA

    L Magnenat, L J Schwimmer & C F Barbas III

  3. Department of Chemistry, The Scripps Research Institute, La Jolla, CA, USA

    L Magnenat, L J Schwimmer & C F Barbas III

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  1. L Magnenat
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Correspondence to C F Barbas III.

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Magnenat, L., Schwimmer, L. & Barbas, C. Drug-inducible and simultaneous regulation of endogenous genes by single-chain nuclear receptor-based zinc-finger transcription factor gene switches. Gene Ther 15, 1223–1232 (2008). https://doi.org/10.1038/gt.2008.96

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