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An Adeno-Associated Virus Vector Efficiently and Specifically Transduces Mouse Skeletal Muscle

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Abstract

Expression of a therapeutic gene in the skeletal muscle is a practical strategy to compensate a patients’ insufficient circulating factor. Its clinical application requires a muscle-targeting vector capable of inducing a continuous high-level transgene expression. We modified an adeno-associated virus serotype 2 (AAV2) vector expressing luciferase from the mouse muscle creatine kinase gene promoter-enhancer (Ckm). First, AAVS1 insulator was inserted into the vector genome for transcriptional enhancement. This increased transduction of mouse quadriceps muscle by 11-fold at 4 weeks after intramuscular injection. Second, two capsid modifications were combined (21F capsid): incorporation of a segment of AAV1 capsid to produce a hybrid capsid and substitution of a tyrosine with a phenylalanine. Use of 21F capsid increased muscle transduction further by 18-fold, resulting in 200-fold higher efficacy than that of the unmodified vector. Compared with a vector having human elongation factor 1α promoter which showed similar efficacy in the muscle, this vector having Ckm transduced non-muscle organs less efficiently after intravenous administration. The AAV2 vector composed of the modified genome and capsid provides a backbone to develop a clinical vector expressing a therapeutic gene in the muscle.

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Acknowledgments

We thank Dr. Kunito Yoshiike for critical reading of the manuscript. This work was supported by Health and Labour Sciences Research Grants from the Ministry of Health, Labour and Welfare.

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Authors and Affiliations

  1. Pathogen Genomics Center, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo, 162-8640, Japan

    Isao Murakami, Takamasa Takeuchi, Seiichiro Mori & Tadahito Kanda

  2. Department of Obstetrics and Gynecology, School of Medicine, Keio University, Tokyo, Japan

    Isao Murakami, Takuma Fujii & Daisuke Aoki

  3. Department of Obstetrics and Gynecology, Faculty of Medicine, The University of Tokyo, Tokyo, Japan

    Mayuyo Mori-Uchino

  4. Department of Radiology, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan

    Keiichi Nakagawa

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  1. Isao Murakami
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  2. Takamasa Takeuchi
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  4. Seiichiro Mori
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Correspondence to Takamasa Takeuchi.

Additional information

Isao Murakami and Takamasa Takeuchi contributed equally to this work.

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Murakami, I., Takeuchi, T., Mori-Uchino, M. et al. An Adeno-Associated Virus Vector Efficiently and Specifically Transduces Mouse Skeletal Muscle. Mol Biotechnol 49, 1–10 (2011). https://doi.org/10.1007/s12033-010-9369-z

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