Sites in the AAV5 capsid tolerant to deletions and tandem duplications.
Sites in the AAV5 capsid tolerant to deletions and tandem duplications.
复制标题
DOI：
10.1016/j.abb.2010.01.009
复制
发表时间：
2010-04-01
影响因子：
3.9
通讯作者：
Ostermeier, Marc
中科院分区：
文献类型：
--
作者：
Hida, Kaoru;Won, Sang Y.;Di Pasquale, Giovanni;Hanes, Justin;Chiorini, John A.;Ostermeier, Marc
基于腺相关病毒（AAV）的基因治疗载体在治疗多种疾病的临床试验中显示出巨大的前景。然而，操纵和改造病毒表面以提高效率的能力对于克服目前限制 AAV 应用的现有免疫和非靶细胞转导等障碍是必要的。尽管先前已经探索了选定位点的单个氨基酸变化和肽插入，但 AAV 对序列小缺失和串联重复的耐受性尚未得到全面解决。在这里，我们生成了一个包含超过 105 个成员的大型多样化文库，其中包含整个 AAV5 病毒衣壳的缺失和串联重复。鉴定出四种独特的突变体，它们保持形成病毒颗粒的能力，其中一种突变体在 293T 和 BEAS-2B 细胞上的转导均得到改善。这种方法可能会潜在地用于产生具有改进和改变的特性的新变体，或者用于鉴定耐受靶向配体插入的位点。
Gene therapy vectors based on adeno-associated virus (AAV) have shown much promise in clinical trials for the treatment of a variety of diseases. However, the ability to manipulate and engineer the viral surface for enhanced efficiency is necessary to overcome such barriers as pre-existing immunity and transduction of non-target cells that currently limit AAV applications. Although single amino acid changes and peptide insertions at select sites have been explored previously, the tolerance of AAV to small deletions and tandem duplications of sequence has not been globally addressed. Here, we have generated a large, diverse library of >105 members containing deletions and tandem duplications throughout the viral capsid of AAV5. Four unique mutants were identified that maintain the ability to form viral particles, with one showing improved transduction on both 293T and BEAS-2B cells. This approach may find potential use for the generation of novel variants with improved and altered properties or in the identification of sites that are tolerant to insertions of targeting ligands.
登录
查看更多内容
DOI：
10.1073/pnas.96.7.3562
发表时间：
1999-03-30
影响因子：
11.1
作者：
Ostermeier, M;Nixon, AE;Benkovic, SJ
通讯作者：
Benkovic, SJ
影响因子：
20.3
作者：
Niemeyer, Glenn P.;Herzog, Roland W.;Lothrop, Clinton D., Jr.
通讯作者：
Lothrop, Clinton D., Jr.
影响因子：
12.4
作者：
Liu, YH;Okada, T;Ozawa, K
通讯作者：
Ozawa, K
影响因子：
12.4
作者：
Koerber, James T.;Jang, Jae-Hyung;Schaffer, David V.
通讯作者：
Schaffer, David V.
影响因子：
5.1
作者：
Sumner-Jones, S. G.;Davies, L. A.;Hyde, S. C.
通讯作者：
Hyde, S. C.