Mandell Jeffrey G, Barbas Carlos F
Department of Molecular Biology and The Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, CA 92037, USA.
Nucleic Acids Res. 2006 Jul 1;34(Web Server issue):W516-23. doi: 10.1093/nar/gkl209.
Individual zinc finger (ZF) domains that recognize DNA triplets with high specificity and affinity can be used to create designer transcription factors and nucleases that are specific for nearly any site in the genome. These domains can be treated as modular units and assembled to create a polydactyl protein that recognizes extended DNA sequences. Deter-mination of valid target sites and the subsequent design of ZF proteins (ZFPs) is error-prone and not trivial, however. As a result, the use of ZFPs have been restricted primarily to those labs with the appropriate expertise. To address these limitations, we have created a user-friendly utility called Zinc Finger Tools (ZF Tools) that can be accessed at the URL http://www.zincfingertools.org. User-supplied DNA sequences can be searched for target sites appropriate for either gene regulation or nuclease targeting. Using a database of experimentally characterized zinc finger domains, the amino acid sequence for a ZFP expected to bind to any chosen target site can be generated. A reverse engineering utility is provided to predict the binding site for a ZFP of known sequence.
能够以高特异性和亲和力识别DNA三联体的单个锌指(ZF)结构域,可用于创建对基因组中几乎任何位点都具有特异性的定制转录因子和核酸酶。这些结构域可被视为模块化单元,并组装成能识别延伸DNA序列的多锌指蛋白。然而,确定有效的靶位点以及随后设计锌指蛋白(ZFPs)容易出错且并非易事。因此,锌指蛋白的使用主要局限于那些具备相应专业知识的实验室。为解决这些限制,我们创建了一个名为“锌指工具”(ZF Tools)的用户友好型实用程序,可通过网址http://www.zincfingertools.org访问。用户提供的DNA序列可用于搜索适合基因调控或核酸酶靶向的靶位点。利用一个经过实验表征的锌指结构域数据库,能够生成预期与任何选定靶位点结合的锌指蛋白的氨基酸序列。还提供了一个逆向工程实用程序,以预测已知序列的锌指蛋白的结合位点。