Beltran Adriana, Liu Yanzheng, Parikh Shaunak, Temple Brenda, Blancafort Pilar
Department of Pharmacology, University of North Carolina, Chapel Hill, NC 27599, USA.
Assay Drug Dev Technol. 2006 Jun;4(3):317-31. doi: 10.1089/adt.2006.4.317.
Artificial transcription factors (ATFs) are proteins designed to specifically bind and regulate genes. Because of their DNA-binding selectivity and modular organization, arrays of zinc finger (ZF) domains have traditionally been used to build the ATF's DNA-binding domains. ATFs have been designed and constructed to regulate a variety of therapeutic targets. Recently, novel combinatorial technologies have been developed to induce expression of any gene of interest or to modify cellular phenotypes. Large repertoires of ATFs have been generated by recombination of all available sequence-specific ZF lexicons. These libraries comprise millions of ATFs with unique DNA-binding specificities. The ATFs are produced by combinatorial assembly of three- and six-ZF building blocks and are linked to activator or repressor domains. Upon delivery into a cell population, any gene in the human genome can potentially be regulated. ATF library members generate genome-wide, experimental perturbations of gene expression, resulting in a phenotypically diverse population, or cellular library. A variety of phenotypic screenings can be applied to select for cells exhibiting a phenotype of interest. The ATFs are then used as genetic probes to identify the targeted genes responsible for the phenotypic switch. In this review we will summarize several applications of ATF library screenings in gene discovery, biotechnology, and disease therapeutics.
人工转录因子（ATF）是经设计可特异性结合并调控基因的蛋白质。由于其DNA结合选择性和模块化结构，传统上一直使用锌指（ZF）结构域阵列构建ATF的DNA结合结构域。已设计并构建出可调控多种治疗靶点的ATF。最近，已开发出新型组合技术来诱导任何感兴趣基因的表达或改变细胞表型。通过重组所有可用的序列特异性ZF词汇表，已产生了大量的ATF。这些文库包含数百万具有独特DNA结合特异性的ATF。ATF由三锌指和六锌指构建模块组合组装而成，并与激活域或抑制域相连。将其导入细胞群体后，人类基因组中的任何基因都有可能受到调控。ATF文库成员会在全基因组范围内对基因表达进行实验性干扰，从而产生表型多样的群体，即细胞文库。可应用多种表型筛选方法来选择表现出感兴趣表型的细胞。然后将ATF用作遗传探针来鉴定导致表型转换的靶向基因。在本综述中，我们将总结ATF文库筛选在基因发现、生物技术和疾病治疗中的几种应用。
