Rapid and highly efficient transduction by double-stranded adeno-associated virus vectors in vitro and in vivo
Rapid and highly efficient transduction by double-stranded adeno-associated virus vectors in vitro and in vivo
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DOI：
10.1038/sj.gt.3302133
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发表时间：
2003-12-01
期刊：
影响因子：
5.1
通讯作者：
Xiao, X
中科院分区：
文献类型：
--
作者：
Wang, Z;Ma, HI;Xiao, X
腺相关病毒（Adeno-associated virus， AAV）是一种基于单链DNA病毒的极具发展前景的基因载体。它的转基因表达需要将ssDNA转化为双链基因组，这是一个缓慢的过程，导致转导延迟和偶尔的低效率。通过突变反向末端重复序列，我们获得了新的AAV载体，主要包装自互补的dsDNA基因组。dsAAV在体外和体内均表现出优越和加速的转导。在大多数检测的细胞系中观察到转基因表达的急剧增加，包括B16黑色素瘤和3LL肺癌，这些细胞系很难被传统的ssAAV载体转导。在包括肌肉和肝脏在内的多种组织中也观察到类似的增加。dsAAV转导绝大多数肝细胞超过6个月，而ssAAV仅转导一小部分。除了避开DNA合成的需要外，dsAAV在体内表现出更高的DNA稳定性和更有效的循环化，这为更快、更强、更持久的转基因表达提供了潜在的分子机制。
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.