Tan Siyuan, Guschin Dmitry, Davalos Albert, Lee Ya-Li, Snowden Andrew W, Jouvenot Yann, Zhang H Steven, Howes Katherine, McNamara Andrew R, Lai Albert, Ullman Chris, Reynolds Lindsey, Moore Michael, Isalan Mark, Berg Lutz-Peter, Campos Bradley, Qi Hong, Spratt S Kaye, Case Casey C, Pabo Carl O, Campisi Judith, Gregory Philip D
Sangamo BioSciences, Inc., Point Richmond Tech Center II, 501 Canal Boulevard, Richmond, CA 94804, USA.
Proc Natl Acad Sci U S A. 2003 Oct 14;100(21):11997-2002. doi: 10.1073/pnas.2035056100. Epub 2003 Sep 26.
Zinc-finger protein transcription factors (ZFP TFs) can be designed to control the expression of any desired target gene, and thus provide potential therapeutic tools for the study and treatment of disease. Here we report that a ZFP TF can repress target gene expression with single-gene specificity within the human genome. A ZFP TF repressor that binds an 18-bp recognition sequence within the promoter of the endogenous CHK2 gene gives a >10-fold reduction in CHK2 mRNA and protein. This level of repression was sufficient to generate a functional phenotype, as demonstrated by the loss of DNA damage-induced CHK2-dependent p53 phosphorylation. We determined the specificity of repression by using DNA microarrays and found that the ZFP TF repressed a single gene (CHK2) within the monitored genome in two different cell types. These data demonstrate the utility of ZFP TFs as precise tools for target validation, and highlight their potential as clinical therapeutics.
锌指蛋白转录因子（ZFP TFs）能够被设计用于控制任何期望的靶基因的表达，从而为疾病的研究和治疗提供潜在的治疗工具。在此我们报告，一种ZFP TF能够在人类基因组内以单基因特异性抑制靶基因表达。一种结合内源性CHK2基因启动子内一个18碱基对识别序列的ZFP TF阻遏物，使CHK2 mRNA和蛋白质水平降低超过10倍。这种抑制水平足以产生功能性表型，DNA损伤诱导的CHK2依赖性p53磷酸化的缺失证明了这一点。我们通过使用DNA微阵列确定了抑制的特异性，发现该ZFP TF在两种不同细胞类型中抑制了监测基因组内的单个基因（CHK2）。这些数据证明了ZFP TFs作为用于靶标验证的精确工具的实用性，并突出了它们作为临床治疗药物的潜力。
