Rapid and highly efficient transduction by double-stranded adeno-associated virus vectors in vitro and in vivo

被引:322
作者
Wang, Z
Ma, HI
Li, J
Sun, L
Zhang, J
Xiao, X
机构
[1] Univ Pittsburgh, Sch Med, Dept Mol Genet & Biochem, Pittsburgh, PA 15261 USA
[2] Univ Pittsburgh, Sch Med, Dept Orthoped Surg, Pittsburgh, PA 15261 USA
[3] Tri Serv Gen Hosp, Natl Def Med Ctr, Dept Neurol Surg, Taipei, Taiwan
关键词
AAV; gene therapy; vector; double-stranded; dsAAV;
D O I
10.1038/sj.gt.3302133
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Adeno-associated virus (AAV) is a promising gene vector based on a single-stranded (ss) DNA virus. Its transgene expression requires the conversion of ssDNA to double-stranded (ds) genome, a slow process responsible for the delayed transduction and occasional inefficiency. By mutating the inverted terminal repeat, we have made novel AAV vectors that predominantly package the self-complementary dsDNA genome. The dsAAV consistently demonstrated superior and accelerated transduction in vitro and in vivo. Dramatic increases in transgene expression were observed in most of the cell lines examined, including B16 melanoma and 3LL lung cancer that are difficult to be transduced by the conventional ssAAV vectors. Similar increases were also observed in vivo in a variety of tissues including muscle and liver. The dsAAV transduced a vast majority of the hepatocytes for more than 6 months, while the ssAAV transduced only a small fraction. In addition to circumventing the requirement for DNA synthesis, the dsAAV exhibited higher in vivo DNA stability and more effective circularization than the ssAAV, suggesting potential molecular mechanisms for the faster, stronger and prolonged transgene expression.
引用
收藏
页码:2105 / 2111
页数:7
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