TY - JOUR
T1 - An adeno-associated virus vector efficiently and specifically transduces mouse skeletal muscle
AU - Murakami, Isao
AU - Takeuchi, Takamasa
AU - Mori-Uchino, Mayuyo
AU - Mori, Seiichiro
AU - Fujii, Takuma
AU - Aoki, Daisuke
AU - Nakagawa, Keiichi
AU - Kanda, Tadahito
N1 - Funding Information:
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.
PY - 2011/9
Y1 - 2011/9
N2 - 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.
AB - 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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U2 - 10.1007/s12033-010-9369-z
DO - 10.1007/s12033-010-9369-z
M3 - Article
C2 - 21197588
AN - SCOPUS:79960951853
SN - 1073-6085
VL - 49
SP - 1
EP - 10
JO - Molecular Biotechnology
JF - Molecular Biotechnology
IS - 1
ER -