Isolation of targeted AAV2 vectors from novel virus display libraries
Isolation of targeted AAV2 vectors from novel virus display libraries
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从新型病毒展示库中分离目标 AAV2 载体
DOI：
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发表时间：
2006
影响因子：
3.5
通讯作者：
J. Kleinschmidt
中科院分区：
文献类型：
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作者：
Daniel A. Waterkamp;O. Müller;Y. Ying;M. Trepel;J. Kleinschmidt
病毒衣壳上展示的随机肽配体是用于选择目标基因转移载体的新兴工具，即使事先不了解潜在的靶细胞受体。我们之前引入了腺相关病毒 (AAV) 展示肽库，确保包装的 AAV 基因组对展示的肽进行编码。这些文库的一个主要限制是它们受到野生型 (wt) AAV 的污染。在这里，我们描述了一种新颖且改进的文库生产系统，该系统通过使用合成的 cap 基因可靠地避免了 wt AAV 的生成。在具有不同wt AAV2复制允许性的细胞类型上，从含wt文库和新型无wt文库中选择靶向AAV载体表明了无wt文库的优越性。然而，从两个文库中选择了高度特异性的肽序列基序，这改善了对 AAV2 复制具有中等或低容许度的细胞的转导。与wt AAV2相比，HeLa细胞转导明显减少，并且一些选定的克隆仅低水平转导非靶细胞，这表明基因转移的效率和特异性均得到提高。总之，我们的研究验证并提高了病毒展示文库在开发靶向基因转移载体方面的独特潜力。版权所有 © 2006 约翰·威利父子有限公司
Random peptide ligands displayed on viral capsids are emerging tools for selection of targeted gene transfer vectors even without prior knowledge of the potential target cell receptor. We have previously introduced adeno‐associated viral (AAV)‐displayed peptide libraries that ensure encoding of displayed peptides by the packaged AAV genome. A major limitation of these libraries is their contamination with wild‐type (wt) AAV. Here we describe a novel and improved library production system that reliably avoids generation of wt AAV by use of a synthetic cap gene. Selection of targeted AAV vectors from wt‐containing and the novel wt‐free libraries on cell types with different permissivity for wt AAV2 replication suggested the superiority of the wt‐free library. However, from both libraries highly specific peptide sequence motifs were selected which improved transduction of cells with moderate or low permissivity for AAV2 replication. Strong reduction of HeLa cell transduction compared to wt AAV2 and only low level transduction of non‐target cells by some selected clones showed that not only the efficiency but also the specificity of gene transfer was improved. In conclusion, our study validates and improves the unique potential of virus display libraries for the development of targeted gene transfer vectors. Copyright © 2006 John Wiley & Sons, Ltd.
影响因子：
4.2
作者：
Shi, WF;Arnold, GS;Bartlett, JS
通讯作者：
Bartlett, JS