Papworth Monika, Kolasinska Paulina, Minczuk Michal
MRC Laboratory of Molecular Biology, Hills Road, CB2 2QH, UK.
Gene. 2006 Jan 17;366(1):27-38. doi: 10.1016/j.gene.2005.09.011. Epub 2005 Nov 17.
The Cys(2)His(2) zinc finger is one of the most common DNA-binding motifs in Eukaryota. A simple mode of DNA recognition by the Cys(2)His(2) zinc finger domain provides an ideal scaffold for designing proteins with novel sequence specificities. The ability to bind specifically to virtually any DNA sequence combined with the potential of fusing them with effector domains has led to the technology of engineering of chimeric DNA-modifying enzymes and transcription factors. This in turn has opened the possibility of using the engineered zinc finger-based factors as novel human therapeutics. One such synthetic factor-designer zinc finger transcription activator of the vascular endothelial growth factor A gene-has recently entered clinical trials to evaluate the ability of stimulating the growth of blood vessels in treating the peripheral arterial obstructive disease. This review concentrates on the aspects of natural Cys(2)His(2) zinc fingers evolution and fundamental steps in design of engineered zinc finger proteins. The applications of engineered zinc finger proteins are discussed in a context of the mechanism mediating their effect on the targeted DNA. Furthermore, the regulation of the expression of zinc finger proteins and their targeting to various cellular compartments and to chromatin and non-chromatin target templates are described. Also possible future applications of designer zinc finger proteins are discussed.
Cys(2)His(2)锌指是真核生物中最常见的DNA结合基序之一。Cys(2)His(2)锌指结构域识别DNA的简单模式为设计具有新序列特异性的蛋白质提供了理想的支架。能够特异性结合几乎任何DNA序列，并具有与效应结构域融合的潜力，这催生了嵌合DNA修饰酶和转录因子的工程技术。这反过来又开启了将工程化的基于锌指的因子用作新型人类治疗药物的可能性。一种这样的合成因子——血管内皮生长因子A基因的设计锌指转录激活剂——最近已进入临床试验，以评估其在治疗外周动脉阻塞性疾病中刺激血管生长的能力。本综述着重于天然Cys(2)His(2)锌指的进化方面以及工程化锌指蛋白设计的基本步骤。在介导其对靶向DNA作用的机制背景下讨论了工程化锌指蛋白的应用。此外，还描述了锌指蛋白表达的调控及其靶向各种细胞区室、染色质和非染色质靶模板的情况。同时也讨论了设计锌指蛋白未来可能的应用。
