Mamounas M, Leavitt M, Yu M, Wong-Staal F
Department of Medicine, University of California, San Diego, La Jolla 92093-0665, USA.
Gene Ther. 1995 Aug;2(6):429-32.
One of the principal problems with using recombinant adeno-associated virus vectors (rAAV) as vehicles for gene delivery has been the difficulty in obtaining high-titer virus stocks after the initial transfection into producer cells. In this report we describe a method for transfecting cells at extremely high efficiency with rAAV vector DNA and complementation plasmid while simultaneously infecting those cells with replication competent adenovirus using adenovirus-polylysine-DNA complexes. We further show that this technique results in an increase in rAAV transducing titer by two orders of magnitude over what is typically achieved by standard calcium phosphate transfection.
将重组腺相关病毒载体（rAAV）用作基因递送载体的主要问题之一是，在最初转染到生产细胞后，难以获得高滴度的病毒储备。在本报告中，我们描述了一种方法，即使用腺病毒-聚赖氨酸-DNA复合物，以极高的效率用rAAV载体DNA和互补质粒转染细胞，同时用具有复制能力的腺病毒感染这些细胞。我们进一步表明，与标准磷酸钙转染通常实现的滴度相比，该技术可使rAAV转导滴度提高两个数量级。
