Strength and muscle specificity of a compact promoter derived from the slow troponin I gene in the context of episomal (gutless adenovirus) and integrating (lentiviral) vectors

Authors

  • Marc-André Robert,

    1. Biotechnology Research Institute, National Research Council Canada, Montreal, Québec, Canada
    2. Programmes de Biologie Moléculaire, Université de Montréal, Montréal, Québec, Canada
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  • Yuanbang Lin,

    1. Department of General Surgery, Tianjin Medical University General Hospital, Tianjin, Heping District, China
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  • Mehdi Bendjelloul,

    1. Biotechnology Research Institute, National Research Council Canada, Montreal, Québec, Canada
    2. Neuromuscular Research Group, Montreal Neurological Institute, McGill University, Montréal, Québec, Canada
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  • Yue Zeng,

    1. Biotechnology Research Institute, National Research Council Canada, Montreal, Québec, Canada
    2. Neuromuscular Research Group, Montreal Neurological Institute, McGill University, Montréal, Québec, Canada
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  • Sofien Dessolin,

    1. Biotechnology Research Institute, National Research Council Canada, Montreal, Québec, Canada
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  • Sophie Broussau,

    1. Biotechnology Research Institute, National Research Council Canada, Montreal, Québec, Canada
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  • Nancy Larochelle,

    1. Neuromuscular Research Group, Montreal Neurological Institute, McGill University, Montréal, Québec, Canada
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  • Josephine Nalbantoglu,

    1. Neuromuscular Research Group, Montreal Neurological Institute, McGill University, Montréal, Québec, Canada
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  • Bernard Massie,

    1. Biotechnology Research Institute, National Research Council Canada, Montreal, Québec, Canada
    2. Département de Microbiologie et Immunologie, Université de Montréal, Montréal, Québec, Canada
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  • Rénald Gilbert

    Corresponding author
    1. Neuromuscular Research Group, Montreal Neurological Institute, McGill University, Montréal, Québec, Canada
    • Biotechnology Research Institute, National Research Council Canada, Montreal, Québec, Canada
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R. Gilbert, Biotechnology Research Institute, 6100 Royalmount Avenue, Montréal, Québec, H4P 2R2, Canada.

E-mail: renald gilbert@cnrc-nrc.gc.ca

Abstract

Background

Gutless adenovirus (helper-dependent adenoviral vector; HDAd) and lentiviral vectors (LV) are attractive vectors for the gene therapy of muscle diseases. Because the organization of their DNA (episomal versus integrated) differs, we investigated whether the strength and specificity of ΔUSEx3, a novel muscle-specific promoter previously tested with plasmid, were maintained in the context of these vectors.

Methods

Two HDAds expressing β-galactosidase regulated by ΔUSEx3 or CAG [cytomegalovirus (CMV) enhancer/β-actin promoter], and three LV expressing green fluorescent protein regulated by ΔUSEx3, CMV or a modified skeletal α-actin promoter (SPcΔ5-12), were constructed. Gene expression was compared in cell culture and after intravenous (HDAd only) and intramuscular injection of mice.

Results

Irrespective of the vector used, ΔUSEx3 remained poorly active in nonmuscle cells and tissues. In myotubes, ΔUSEx3 was as strong as CMV and SPcΔ5-12, although it was ten-fold weaker than CAG, a proven powerful promoter in muscle. In cell culture, ΔUSEx3 activity in the context of LV was more stable than CMV, indicating it is less prone to silencing. In the context of HDAd, the behavior of ΔUSEx3 in skeletal muscle mirrored that of cell culture (10% of the CAG activity and half the number of transduced fibers). Surprisingly, in muscles treated with LV, ΔUSEx3 activity was five-fold lower than SPcΔ5-12.

Conclusions

The data obtained in the present study confirm that ΔUSEx3 is a strong and robust muscle-specific promoter in the context of HDAd (cell culture and in vivo) and LV (cell culture). However, it was less efficient in vivo in the context of LV.

Copyright © 2012 National Research Council Canada & John Wiley & Sons, Ltd.

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