Positive and negative regulation of endogenous genes by designed transcription factors
Positive and negative regulation of endogenous genes by designed transcription factors
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DOI：
10.1073/pnas.040552697
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发表时间：
2000-02-15
影响因子：
11.1
通讯作者：
Barbas, CF
中科院分区：
文献类型：
--
作者：
Beerli, RR;Dreier, B;Barbas, CF
通过利用设计的锌指蛋白研究了强制定位的基因调控，这些锌指蛋白结合原癌基因erbB - 2和erbB - 3的5'非翻译区的18bp DNA序列。通过将DNA结合蛋白与抑制或激活结构域融合产生转录因子。当将这些转录因子引入细胞时，它们分别作为内源性erbB - 2或erbB - 3基因表达的显性抑制因子或激活因子起作用。重要的是，尽管在erbB - 2和erbB - 3中靶向的转录因子结合位点有18个核苷酸中的15个是相同的，但对这两个基因的强制调控具有高度特异性。在保留DNA靶序列的几个物种来源的细胞中观察到了对erbB - 2基因表达的调控。在SKBR3乳腺癌细胞中抑制erbB - 2可通过诱导G₁期积累来抑制细胞周期进程，这表明设计的转录因子在癌症基因治疗中的潜力。这些结果证明了内源性基因可被有意地向上和向下调控，并提供了一种改变生物系统的额外手段。
Gene regulation by imposed localization was studied by using designed zinc finger proteins that bind 18-bp DNA sequences in the 5' untranslated regions of the protooncogenes erbB-2 and erbB-3, Transcription factors were generated by fusion of the DNA-binding proteins to repression or activation domains, When introduced into cells these transcription factors acted as dominant repressors or activators of, respectively, endogenous erbB-2 or erbB-3 gene expression. significantly, imposed regulation of the two genes was highly specific, despite the fact that the transcription factor binding sites targeted in erbB-2 and erbB-3 share 15 of 18 nucleotides. Regulation of erbB-2 gene expression was observed in cells derived from several species that conserve the DNA target sequence. Repression of erbB-2 in SKBR3 breast cancer cells inhibited cell-cycle progression by inducing a G(1) accumulation, suggesting the potential of designed transcription factors for cancer gene therapy. These results demonstrate the willful up- and down-regulation of endogenous genes, and provide an additional means to alter biological systems.