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Delivering multiple gene products in the brain from a single adeno-associated virus vector

Gene Therapy volume 16, pages 1314–1319 (2009)Cite this article

Abstract

For certain gene therapy applications, the simultaneous delivery of multiple genes would allow for novel therapies. In the case of adeno-associated virus (AAV) vectors, the limited packaging capacity greatly restricts current methods of carrying multiple transgene cassettes. To address this issue, a recombinant AAV (rAAV) vector was designed such that a furin proteolytic cleavage site (RKRRKR) was placed between the coding sequences of two genes (green fluorescent protein (GFP) and galanin), to allow cleavage of the chimeric protein into two fragments. In addition, these constructs contained the fibronectin secretory signal sequence that causes the gene products to be constitutively secreted from transduced cells. In vitro studies show that after transfection of HEK293 cells, the appropriate cleavage and constitutive secretion occurred regardless of the order of the genes in the transgene cassette. In vivo, infusion of rAAV vectors into the piriform cortex resulted in both GFP expression and significant galanin attenuation of kainic acid-induced seizure activity. Thus, the present results establish the utility of a proteolytic approach for the expression and secretion of multiple gene products from a single AAV vector transgene cassette.

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Acknowledgements

These studies were supported by NINDS Grant NS 35633 to TJM.

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Authors and Affiliations

  1. Gene Therapy Center, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA

    S B Foti, R J Samulski & T J McCown

  2. Curriculum in Neurobiology, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA

    S B Foti

  3. Department of Pharmacology, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA

    R J Samulski

  4. Department of Psychiatry, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA

    T J McCown

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  1. S B Foti
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  2. R J Samulski
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Correspondence to T J McCown.

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Foti, S., Samulski, R. & McCown, T. Delivering multiple gene products in the brain from a single adeno-associated virus vector. Gene Ther 16, 1314–1319 (2009). https://doi.org/10.1038/gt.2009.106

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