Rapid "open-source" engineering of customized zinc-finger nucleases for highly efficient gene modification.
Rapid "open-source" engineering of customized zinc-finger nucleases for highly efficient gene modification.
复制标题
DOI：
10.1016/j.molcel.2008.06.016
复制
发表时间：
2008-07-25
期刊：
影响因子：
16
通讯作者：
Joung JK
中科院分区：
文献类型：
--
作者：
Maeder ML;Thibodeau-Beganny S;Osiak A;Wright DA;Anthony RM;Eichtinger M;Jiang T;Foley JE;Winfrey RJ;Townsend JA;Unger-Wallace E;Sander JD;Müller-Lerch F;Fu F;Pearlberg J;Göbel C;Dassie JP;Pruett-Miller SM;Porteus MH;Sgroi DC;Iafrate AJ;Dobbs D;McCray PB Jr;Cathomen T;Voytas DF;Joung JK
定制的锌指核酸酶（ZFN）可以在果蝇、C.线虫植物和人类ZFN技术应用中的瓶颈一直是高度特异性工程化锌指阵列的产生。在这里，我们描述了开放（寡聚池工程师），一个快速的，公开可用的策略，用于构建多指阵列，我们表明是更有效的比以前发表的模块化组装方法。我们使用OPEN构建了37个高活性ZFN对，其在位于三个内源性人类基因（VEGF-A，HoxB 13，CFTR），一个内源性植物基因（烟草SuRA）和一个染色体整合的EGFP报告基因内的11个不同靶位点以高效率（1至50%）诱导靶向改变。总之，OPEN提供了一种快速设计高活性锌指阵列的“开源”方法，从而使ZFN技术在生物研究和基因治疗中的实践、开发和应用更加广泛。
Custom-made zinc-finger nucleases (ZFNs) can induce targeted genome modifications with high efficiency in cell types including Drosophila, C. elegans, plants, and humans. A bottleneck in the application of ZFN technology has been the generation of highly specific engineered zinc-finger arrays. Here we describe OPEN (Oligomerized Pool ENgineering), a rapid, publicly available strategy for constructing multi-finger arrays, which we show is more effective than the previously published modular assembly method. We used OPEN to construct 37 highly active ZFN pairs which induced targeted alterations with high efficiencies (1 to 50%) at 11 different target sites located within three endogenous human genes (VEGF-A, HoxB13, CFTR), an endogenous plant gene (tobacco SuRA), and a chromosomally-integrated EGFP reporter gene. In summary, OPEN provides an “open-source” method for rapidly engineering highly active zinc-finger arrays, thereby enabling broader practice, development, and application of ZFN technology for biological research and gene therapy.
登录
查看更多内容
影响因子：
46.9
作者：
Isalan, M;Klug, A;Choo, Y
通讯作者：
Choo, Y
影响因子：
12.4
作者：
Alwin, S;Gere, MB;Cathomen, T
通讯作者：
Cathomen, T
DOI：
10.1073/pnas.94.11.5617
发表时间：
1997-05-27
影响因子：
11.1
作者：
Isalan, M;Choo, Y;Klug, A
通讯作者：
Klug, A
影响因子：
56.9
作者：
Bibikova, M;Beumer, K;Carroll, D
通讯作者：
Carroll, D
影响因子：
12.4
作者：
Cornu, Tatjana I.;Thibodeau-Beganny, Stacey;Cathomen, Toni
通讯作者：
Cathomen, Toni