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Dual transgene expression by foamy virus vectors carrying an endogenous bidirectional promoter

Gene Therapy volume 17, pages 380–388 (2010)Cite this article

Abstract

Several gene therapy applications require the transfer and simultaneous expression of multiple genes in the same cell. In this study, we analyzed the potential for coordinated expression of an endogenous bidirectional promoter located on chromosome X, which controls the expression of the heterogeneous nuclear ribonucleoprotein H2 (HNRNPH2) and alpha-galactosidase (GLA) genes. The promoter was cloned in both transcriptional orientations in a foamy virus (FV) vector backbone, whereas the enhanced green fluorescent protein (EGFP) and low-affinity nerve growth factor receptor (ΔLNGFR) reporter genes were cloned in the 5′–3′ and 3′–5′ transcriptional orientations, respectively. In all the cell lines tested, both vectors showed high levels of transgene coexpression that reached 76% of total positive cells (range from 76 to 18%). Comparison of EGFP and ΔNGFR levels revealed that the side of the promoter that drives the expression of the HNRNPH2 gene in the genome was stronger and in accordance to its in situ activity. When tested with CD34+ cells, transgene coexpression reached 35.3% of all positive cells in progenitor assays and 16.8% of all positive cells after transplantation in NOD/severe combined immunodeficient mice. In summary, we show that the endogenous promoter used in this study holds bidirectional activity in the context of FV vectors and can be used in gene therapy applications requiring synchronized expression of two genes.

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Acknowledgements

This work was supported in part by the European IP CONSERT (FP6, 005242) and an ENTER Grant to ES (GSRT, Number 04ER022).

Author information

Author notes

  1. A Andrianaki and E K Siapati: These authors contributed equally to this work.

Authors and Affiliations

  1. Cell and Gene Therapy Laboratory, Center for Basic Research II, Biomedical Research Foundation of the Academy of Athens, Athens, Greece

    A Andrianaki, E K Siapati & G Vassilopoulos

  2. Division of Haematology, Department of Medicine, University of Washington, Seattle, WA, USA

    R K Hirata & D W Russell

  3. Division of Haematology, University of Thessaly Medical School, Larissa, Greece

    G Vassilopoulos

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  1. A Andrianaki
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  2. E K Siapati
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  5. G Vassilopoulos
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Correspondence to G Vassilopoulos.

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Andrianaki, A., Siapati, E., Hirata, R. et al. Dual transgene expression by foamy virus vectors carrying an endogenous bidirectional promoter. Gene Ther 17, 380–388 (2010). https://doi.org/10.1038/gt.2009.147

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Keywords

  • FV
  • bidirectional promoter
  • gene transfer
  • hematopoietic stem cells

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