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Evaluation of macrophage-specific promoters using lentiviral delivery in mice

Gene Therapy volume 19, pages 1041–1047 (2012)Cite this article

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Abstract

In gene therapy, tissue-specific promoters are useful tools to direct transgene expression and improve efficiency and safety. Macrophage-specific promoters (MSPs) have previously been published using different delivery systems. In this study, we evaluated five different MSPs fused with green fluorescent protein (GFP) to delineate the one with highest specificity using lentiviral delivery. We compared three variants of the CD68 promoter (full length, the 343-bp proximal part and the 150-bp proximal part) and two variants (in forward and reverse orientation) of a previously characterized synthetic promoter derived from elements of transcription factor genes. We transduced a number of cell lines and primary cells in vitro. In addition, hematopoietic stem cells were transduced with MSPs and transferred into lethally irradiated recipient mice. Fluorescence activated cell sorting analysis was performed to determine the GFP expression in different cell populations both in vitro and in vivo. We showed that MSPs can efficiently be used for lentiviral gene delivery and that the 150-bp proximal part of the CD68 promoter provides primarily macrophage-specific expression of GFP. We propose that this is the best currently available MSP to use for directing transgene expression to macrophage populations in vivo using lentiviral vectors.

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Acknowledgements

We thank Louise Henningsson, Kristina Skålén, Elin Stenfeldt and Maria Heyden (Gothenburg University) for expert technical assistance; Yan Ru Su and John Blakemore (Vanderbilt University) for stimulating discussions and technical expertise; and Rosie Perkins (Gothenburg University) for editing the manuscript. This work was supported by the Swedish Research Council; the VINNOVA Foundation; the IngaBritt and Arne Lundgren Foundation; the Swedish Foundation for Strategic Research; the Sahlgrenska University Hospital ALF research grants; and NIH Grants HL105375, HL57986, HL106845 and HL65709.

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Author notes

  1. J Borén and I Gjertsson: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Molecular and Clinical Medicine,

    M C Levin, U Lidberg, M Adiels, A Wramstedt, S-O Olofsson & J Borén

  2. Wallenberg Laboratory, University of Gothenburg, Göteborg, Sweden

    M C Levin, U Lidberg, M Adiels, A Wramstedt, S-O Olofsson & J Borén

  3. Department of Rheumatology and Inflammation Research, University of Gothenburg, Göteborg, Sweden

    P Jirholt & I Gjertsson

  4. Molecular Immunology Unit, Institute of Child Health, University College London, London, UK

    K Gustafsson

  5. Sir William Dunn School of Pathology, University of Oxford, Oxford, UK

    D R Greaves

  6. Department of Medicine, Audie L Murphy Division, University of Texas Health Science Center at San Antonio, and South Texas Veterans Health Care System, San Antonio, TX, USA

    S Li

  7. Division of Cardiovascular Medicine, Vanderbilt University Medical Center, Nashville, TN, USA

    S Fazio & M F Linton

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  1. M C Levin
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  8. S Li
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Correspondence to M C Levin.

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Levin, M., Lidberg, U., Jirholt, P. et al. Evaluation of macrophage-specific promoters using lentiviral delivery in mice. Gene Ther 19, 1041–1047 (2012). https://doi.org/10.1038/gt.2011.195

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