Zhang X, Li C Y
Department of Radiation Oncology, Duke University Medical Center, Durham, NC 27710, USA.
Mol Ther. 2001 May;3(5 Pt 1):787-92. doi: 10.1006/mthe.2001.0306.
An efficient approach to the large-scale production of rAAV will significantly facilitate the application of this promising gene delivery vector in human gene therapy applications. In this study, we report a novel approach to rAAV production that is based exclusively on the adenovirus vector, without the need for plasmid transfections and special packaging cell lines. All components required for rAAV production, including the rep and cap genes, and the therapeutic gene(s) are delivered to the widely used 293 packaging cell line by adenovirus vectors. High-titer rAAV vectors (200-600 infectious units/producer cell) were obtained by use of this approach. As adenovirus vectors can be produced to high titers and they can infect cells in suspension efficiently, this approach may be amenable to scaled-up production of rAAV vectors.
一种高效生产重组腺相关病毒(rAAV)的方法将显著促进这种有前景的基因递送载体在人类基因治疗中的应用。在本研究中,我们报告了一种全新的rAAV生产方法,该方法完全基于腺病毒载体,无需质粒转染和特殊的包装细胞系。rAAV生产所需的所有组件,包括rep和cap基因以及治疗性基因,均通过腺病毒载体递送至广泛使用的293包装细胞系。通过这种方法获得了高滴度的rAAV载体(每产生细胞200 - 600个感染单位)。由于腺病毒载体能够产生高滴度且能有效感染悬浮细胞,这种方法可能适用于rAAV载体的大规模生产。