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siRNA silencing of keratinocyte-specific GFP expression in a transgenic mouse skin model

Gene Therapy volume 16, pages 963–972 (2009)Cite this article

Abstract

Small interfering RNAs (siRNAs) can be designed to specifically and potently target and silence a mutant allele, with little or no effect on the corresponding wild-type allele expression, presenting an opportunity for therapeutic intervention. Although several siRNAs have entered clinical trials, the development of siRNA therapeutics as a new drug class will require the development of improved delivery technologies. In this study, a reporter mouse model (transgenic click beetle luciferase/humanized monster green fluorescent protein) was developed to enable the study of siRNA delivery to skin; in this transgenic mouse, green fluorescent protein reporter gene expression is confined to the epidermis. Intradermal injection of siRNAs targeting the reporter gene resulted in marked reduction of green fluorescent protein expression in the localized treatment areas as measured by histology, real-time quantitative polymerase chain reaction and intravital imaging using a dual-axes confocal fluorescence microscope. These results indicate that this transgenic mouse skin model, coupled with in vivo imaging, will be useful for development of efficient and ‘patient-friendly’ siRNA delivery techniques and should facilitate the translation of siRNA-based therapeutics to the clinic for treatment of skin disorders.

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Acknowledgements

This study was funded in part by grants from the National Institutes of Health (R43AR055881, RLK and U54 CA105296-01, CHC). We thank Irwin McLean and Birgitte Lane for critical review of the manuscript, Heini Ilves, Manuel Flores and Maria Fernanda Lara for technical support, and Mohammed Ikram and Mike Philpott for the kind gift of phK14 plasmid. EGG is the recipient of a PC Project fellowship.

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Authors and Affiliations

  1. Molecular Imaging Program at Stanford (MIPS), Stanford University School of Medicine, Stanford, CA, USA

    E Gonzalez-Gonzalez, Q Wang, W Piyawattanametha, T C Doyle & C H Contag

  2. Department of Pediatrics, Stanford University School of Medicine, Stanford, CA, USA

    E Gonzalez-Gonzalez, Q Wang, W Piyawattanametha, T C Doyle & C H Contag

  3. Department of Radiology, Stanford University School of Medicine, Stanford, CA, USA

    C H Contag

  4. Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, CA, USA

    C H Contag

  5. Department of Electrical Engineering, Stanford University, Stanford, CA, USA

    H Ra, W Piyawattanametha, M J Mandella, G S Kino & O Solgaard

  6. TransDerm Inc., Santa Cruz, CA, USA

    R P Hickerson & R L Kaspar

  7. NECTEC, Pathumthani, Thailand

    W Piyawattanametha

  8. Thermo Fisher Scientific, Dharmacon Products, Lafayette, CO, USA

    D Leake

  9. Centre for Cutaneous Research, Cancer Research UK, Institute of Cell and Molecular Science, Barts and the London School of Medicine and Dentistry, Whitechapel, London, UK

    A A Avilion

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  1. E Gonzalez-Gonzalez
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  2. H Ra
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  4. Q Wang
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Corresponding author

Correspondence to R L Kaspar.

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Supplementary Information accompanies the paper on Gene Therapy website (http://www.nature.com/gt)

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Gonzalez-Gonzalez, E., Ra, H., Hickerson, R. et al. siRNA silencing of keratinocyte-specific GFP expression in a transgenic mouse skin model. Gene Ther 16, 963–972 (2009). https://doi.org/10.1038/gt.2009.62

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