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Regulation of endogenous gene expression using small molecule-controlled engineered zinc-finger protein transcription factors

Gene Therapy volume 14, pages 1362–1369 (2007)Cite this article

Abstract

Small-molecule-regulated gene expression offers the promise of titrating the dose and duration of action of DNA-based therapies. To this end, we show that engineered zinc-finger protein transcription factors (ZFP TFs) can be coupled with a drug-inducible regulatory domain to permit small-molecule control of endogenous gene transcription. We constructed a drug-responsive ZFP TF via the fusion of a ZFP DNA-binding domain (DBD) targeting the human VEGF-A gene and an effector domain containing a truncated progesterone receptor ligand-binding domain linked to the NFκB p65 activation domain. Introduction of this engineered ZFP TF into human or murine cells allowed expression of the chromosomal VEGF-A gene to be induced upon addition of mifepristone, a synthetic steroid analog. Mifepristone-dependent VEGF-A induction was rapid, dose-dependent and reversible. Moreover, stable lines expressing the drug-responsive ZFP TF could be maintained in a state of continuous induction for at least 30 days without loss of viability. Potent VEGF-A induction was demonstrated using different engineered ZFP DBDs, thus this approach may represent a general solution to small-molecule regulation of targeted endogenous genes.

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Acknowledgements

We thank Drs Trevor Collingwood, Michael Holmes, Edward Rebar and Sean Brennan for careful reading of the manuscript. We are also grateful to Edward Lanphier for encouragement and support.

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Author notes

  1. C L Dent

    Present address: 2Current address: Medical and Molecular Genetics, Kings College London, 8th Floor Guy's Tower, Guy's Hospital, London SE1 9RT, UK.,

  2. C C Case

    Present address: 3Current address: SanBio Inc., 231 S. Whisman Road, Mountain View, CA 94041, USA.,

Authors and Affiliations

  1. Sangamo BioSciences Inc., Point Richmond Tech Center, Richmond, CA, USA

    C L Dent, G Lau, E A Drake, A Yoon, C C Case & P D Gregory

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Correspondence to P D Gregory.

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Dent, C., Lau, G., Drake, E. et al. Regulation of endogenous gene expression using small molecule-controlled engineered zinc-finger protein transcription factors. Gene Ther 14, 1362–1369 (2007). https://doi.org/10.1038/sj.gt.3302985

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