Production methods for gene transfer vectors based on adeno-associated virus serotypes
Production methods for gene transfer vectors based on adeno-associated virus serotypes
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DOI：
10.1016/s1046-2023(02)00219-0
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发表时间：
2002-10-01
期刊：
影响因子：
4.8
通讯作者：
Grimm, D
中科院分区：
文献类型：
--
作者：
Grimm, D
来源于腺相关病毒scrotype 2（AAV-2）的载体代表了最有前途的人类基因转移工具，因为这些载体对靶细胞既无致病性也无毒性，并且允许基因在多种组织中长期表达。然而，它们在感染许多临床相关的细胞类型方面效率相当低，并且这些载体的转导可能受到在人类群体中高度流行的中和抗体的阻碍。越来越多的研究人员目前将注意力转向AAV的其他五种血清型。尝试开发作为人类基因转移的新型载体，希望克服与腺相关病毒2载体相关的问题。在这里，我描述和讨论的方法，以产生这些替代的AAV载体在组织培养。详细地，两种策略进行了比较，依赖于转染的细胞培养物中的两个或三个质粒，含有AAV载体基因组和编码AAV和腺病毒辅助功能。这些方案中的任一种可用于将重组AAV基因组包装到其自身scrotype的衣壳中（“真实的”scrotype的产生）或将该载体DNA“交叉包装”到衍生自另一种AAV血清型的衣壳中（“假型化”）。由于这些方法仍处于其早期阶段，因此讨论了当前技术的现有限制，并提出了可能的进一步改进。(C)2002年由Elsevier Science（美国）出版。
Vectors derived from adeno-associated virus scrotype 2 (AAV-2) represent a most promising tool Or human gene transfer because these vectors are neither pathogenic nor toxic to the target cell, and allow long-term gene expression in a large variety of tissues. However, they are rather inefficient at infecting a number of clinically relevant cell types, and transduction by these vectors is likely hampered by neutralizing antibodies that are highly prevalent in the human Population, Therefore. in increasing number of researchers are currently turning their attention to the five other serotypes of AAV. to try and develop as novel vectors for human gene transfer, hoping to overcome the problems associated with AAV-2 vectors. Here I describe and discuss the methodology to produce these alternative AAV vectors in tissue culture. In detail, two strategies are compared that rely on transfection of cells in culture with either two or three plasmids, containing the AAV vector genome and encoding AAV and adenoviral helper functions. Either of these protocols can be used to package a recombinant AAV genome into capsids of its own scrotype (generation of "real" scrotypes) or to "cross-package" this vector DNA into capsids derived from another AAV serotype ("pseudotyping"), As these approaches are still in their early stages, the existing limitations of current technology, are discussed and possible further improvements proposed. (C) 2002 Published by Elsevier Science (USA).