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Efficient gene transfer in skeletal muscle with AAV-derived bicistronic vector using the FGF-1 IRES

Gene Therapy volume 15, pages 1090–1098 (2008)Cite this article

Abstract

IRESs (internal ribosome entry sites) are RNA elements behaving as translational enhancers in conditions of global translation blockade. IRESs are also useful in biotechnological applications as they allow expression of several genes from a single mRNA. Up to now, most IRES-containing vectors use the IRES from encephalomyocarditis virus (EMCV), highly active in transiently transfected cells but long and not flexible in its positioning relative to the gene of interest. In contrast, several IRESs identified in cellular mRNAs are short and flexible and may therefore be advantageous in gene transfer vectors such as those derived from the adeno-associated virus (AAV), where the size of the transgene expression cassette is limited. Here, we have tested bicistronic AAV-derived vectors expressing two luciferase genes separated by the EMCV- or fibroblast growth factor 1 (FGF-1) IRES. We demonstrate that the AAV vector with the FGF-1 IRES, when administrated into the mouse muscle, leads to efficient expression of both transgenes with a stable stoechiometry, for at least 120 days. Interestingly, the bicistronic mRNA containing the FGF-1 IRES leads to transgene expression 10 times superior to that observed with EMCV, in vivo. AAV vectors featuring the FGF-1 IRES may thus be advantageous for gene therapy approaches in skeletal muscle involving coexpression of genes of interest.

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Acknowledgements

We are grateful for technical assistance to Y Barreira (IFR31 animal facility), to JJ Maoret (IFR31 platform of molecular biology) and MJ Fouque. This work was supported by grants from Association Française contre les Myopathies (AFM), Fondation de l'Avenir Association pour la Recherche sur le Cancer (ARC), Conseil Régional Midi-Pyrénées, European Commission FP5 (QOL-2000-3.1.2, consortium CONTEXTH contract QLRT-2000-00721) and the French Ministery of Research (decision No 01H0387).

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Author notes

  1. C Le Bec and L Van den Berghe: These authors contributed equally to this work.

Authors and Affiliations

  1. Institut National de la Santé et de la Recherche Médicale (INSERM), U858, Toulouse, France

    A Delluc-Clavières, L Van den Berghe, C Conte & A-C Prats

  2. Team ‘Translational control and gene therapy of vascular and tumoral diseases,’ Institut de Médecine Moléculaire de Rangueil, IFR31, Université Toulouse III Paul Sabatier, Toulouse, France

    A Delluc-Clavières, L Van den Berghe, C Conte & A-C Prats

  3. Généthon, Evry, France

    C Le Bec & V Allo

  4. Institut National de la Santé et de la Recherche Médicale (INSERM), U781, Paris, France

    O Danos

  5. Université Paris 5, Paris, France

    O Danos

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  1. A Delluc-Clavières
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  2. C Le Bec
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Correspondence to A-C Prats.

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Delluc-Clavières, A., Le Bec, C., Van den Berghe, L. et al. Efficient gene transfer in skeletal muscle with AAV-derived bicistronic vector using the FGF-1 IRES. Gene Ther 15, 1090–1098 (2008). https://doi.org/10.1038/gt.2008.49

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