Molecular Pathology Unit and Center for Cancer Research, Massachusetts General Hospital, Charlestown, Massachusetts, USA.
Nat Methods. 2011 Jan;8(1):67-9. doi: 10.1038/nmeth.1542. Epub 2010 Dec 12.
Engineered zinc-finger nucleases (ZFNs) enable targeted genome modification. Here we describe context-dependent assembly (CoDA), a platform for engineering ZFNs using only standard cloning techniques or custom DNA synthesis. Using CoDA-generated ZFNs, we rapidly altered 20 genes in Danio rerio, Arabidopsis thaliana and Glycine max. The simplicity and efficacy of CoDA will enable broad adoption of ZFN technology and make possible large-scale projects focused on multigene pathways or genome-wide alterations.
基因工程锌指核酸酶（ZFNs）可实现靶向基因组修饰。在此，我们描述了一种基于上下文的组装（CoDA）平台，该平台仅使用标准克隆技术或定制 DNA 合成即可对 ZFN 进行工程改造。使用 CoDA 生成的 ZFN，我们快速改变了斑马鱼、拟南芥和大豆中的 20 个基因。CoDA 的简单性和高效性将使 ZFN 技术得到广泛应用，并有可能开展针对多基因途径或全基因组改变的大规模项目。
