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Selective enhancement of gene transfer by steroid-mediated gene delivery

Nature Biotechnology volume 19, pages 1155–1161 (2001)Cite this article

Abstract

The incorporation of transgenes into the host cells' nuclei is problematic using conventional nonviral gene delivery technologies. Here we describe a strategy called steroid-mediated gene delivery (SMGD), which uses steroid receptors as shuttles to facilitate the uptake of transfected DNA into the nucleus. We use glucocorticoid receptors (GRs) as a model system with which to test the principle of SMGD. To this end, we synthesized and tested several bifunctional steroid derivatives, finally focusing on a compound named DR9NP, consisting of a dexamethasone backbone linked to a psoralen moiety using a nine-atom chemical spacer. DR9NP binds to the GR in either its free or DNA-crosslinked form, inducing the translocation of the GR to the nucleus. The expression of transfected DR9NP-decorated reporter plasmids is enhanced in dividing cells: expression of steroid-decorated reporter plasmids depends on the presence of the GR, is independent of the transactivation potential of the GR and correlates with enhanced nuclear accumulation of the transgene in GR-positive cells. The SMGD effect is also observed in cells naturally expressing GRs and is significantly increased in nondividing cell cultures. We propose that SMGD could be used as a platform for selective targeting of transgenes in nonviral somatic gene transfer.

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Acknowledgements

We thank R. Heim and M. Imhof, respectively, for the gift of the GFP cDNA and the expression vector for luciferase. Our thanks go also to B. Huse for the assembly of recombinant adenoviral vectors for GR. The contributions of P. Matthey, P. de los Rios and M. Woodle for assistance, math coaching and critical discussions are also gratefully acknowledged. This work has been supported by the Canton of Fribourg, the Canton of Bern, The Inselspital and in large part by the Swiss National Research Program NFP37 “somatic gene therapy” (http://www.unifr.ch/nfp37).

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Author notes

  1. These three authors have contributed equally to this work.

Authors and Affiliations

  1. Division of Nephrology, Inselspital, Bern, Switzerland

    Alexandre Rebuffat, Alessio Bernasconi, Hans Wehrli, Brigitte M. Frey & Felix J. Frey

  2. Institute of Biochemistry, University of Fribourg (UNIFR), Fribourg, Switzerland

    Maurizio Ceppi, Stefano Brenz Verca & Sandro Rusconi

  3. Institute of Histology, UNIFR, Fribourg, Switzerland

    Merdol Ibrahim

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  1. Alexandre Rebuffat
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The online version of the original article can be found at 10.1038/nbt1201-1118

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Rebuffat, A., Bernasconi, A., Ceppi, M. et al. Selective enhancement of gene transfer by steroid-mediated gene delivery. Nat Biotechnol 19, 1155–1161 (2001). https://doi.org/10.1038/nbt1201-1155

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