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Self-complementary recombinant adeno-associated virus (scAAV) vectors promote efficient transduction independently of DNA synthesis

Gene Therapy volume 8, pages 1248–1254 (2001)Cite this article

Abstract

Adeno-associated virus (AAV) vectors package single-stranded genomes and require host-cell synthesis of the complementary strand for transduction. However, when the genome is half wild-type size, AAV can package either two copies, or dimeric inverted repeat DNA molecules. Dimeric, or self-complementary molecules (scAAV) should spontaneously reanneal, alleviating the requirement for host-cell DNA synthesis. We generated and characterized scAAV vectors in order to bypass the rate-limiting step of second-strand synthesis. In vitro, scAAV vectors were five- to 140-fold more efficient transducing agents than conventional rAAV, with a 5.9:1 particle to transducing unit ratio. This efficiency is neither greatly increased by co-infection with Ad, nor inhibited by hydroxyurea, demonstrating that transduction is independent of DNA synthesis. In vivo, scAAV expressing erythropoietin resulted in rapid and higher levels of hematocrit than a conventional single-stranded vector. These novel scAAV vectors represent a biochemical intermediate in rAAV transduction and should provide new insights into the biology of vector transduction.

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Acknowledgements

We acknowledge the technical assistance of Jennifer Naspinski with scAAV studies in vivo, and Xiaohuai Zhou from the UNC Vector Core for construction of rAAV/epo/lambda vector. This work was supported in part by NIH grants HL 48347, 51818. Doug McCarty was supported in part by NIH grant HL 51818 and PEM receives research support from the National Hemophilia Foundation and from NIH HL 03960.

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Authors and Affiliations

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

    DM McCarty, PE Monahan & RJ Samulski

  2. School of Pharmacy, University of North Carolina at Chapel Hill, NC, USA

    DM McCarty

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

    PE Monahan

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

    RJ Samulski

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  1. DM McCarty
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  2. PE Monahan
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  3. RJ Samulski
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McCarty, D., Monahan, P. & Samulski, R. Self-complementary recombinant adeno-associated virus (scAAV) vectors promote efficient transduction independently of DNA synthesis. Gene Ther 8, 1248–1254 (2001). https://doi.org/10.1038/sj.gt.3301514

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