Department of Chemistry, North Carolina State University, Raleigh, North Carolina 27695, USA.
J Am Chem Soc. 2011 Aug 24;133(33):13176-82. doi: 10.1021/ja204980v. Epub 2011 Jul 29.
DNA decoys have been developed for the inhibition of transcriptional regulation of gene expression. However, the present methodology lacks the spatial and temporal control of gene expression that is commonly found in nature. Here, we report the application of photoremovable protecting groups on nucleobases of nuclear factor κB (NF-κB) DNA decoys to regulate NF-κB-driven transcription of secreted alkaline phosphatase using light as an external control element. The NF-κB family of proteins is comprised of important eukaryotic transcription factors that regulate a wide range of cellular processes and are involved in immune response, development, cellular growth, and cell death. Several diseases, including cancer, arthritis, chronic inflammation, asthma, neurodegenerative diseases, and heart disease, have been linked to constitutively active NF-κB. Through the direct incorporation of caging groups into an NF-κB decoy, we were able to disrupt DNA:DNA hybridization and inhibit the binding of the transcription factor to the DNA decoy until UV irradiation removed the caging groups and restored the activity of the oligonucleotide. Excellent light-switching behavior of transcriptional regulation was observed. This is the first example of a caged DNA decoy for the photochemical regulation of gene expression in mammalian cells and represents an important addition to the toolbox of light-controlled gene regulatory agents.
DNA 诱饵已被开发用于抑制基因表达的转录调控。然而，目前的方法缺乏自然界中常见的基因表达的时空控制。在这里，我们报告了在核因子 κB（NF-κB）DNA 诱饵的核碱基上应用光可去除保护基团，以用光作为外部控制元件来调节 NF-κB 驱动的分泌型碱性磷酸酶的转录。NF-κB 蛋白家族由重要的真核转录因子组成，它们调节广泛的细胞过程，并参与免疫反应、发育、细胞生长和细胞死亡。几种疾病，包括癌症、关节炎、慢性炎症、哮喘、神经退行性疾病和心脏病，都与持续激活的 NF-κB 有关。通过将笼状基团直接掺入 NF-κB 诱饵中，我们能够破坏 DNA:DNA 杂交并抑制转录因子与 DNA 诱饵的结合，直到 UV 照射去除笼状基团并恢复寡核苷酸的活性。观察到转录调控的出色光开关行为。这是第一个用于在哺乳动物细胞中光化学调控基因表达的笼状 DNA 诱饵的例子，是光控基因调节剂工具包的重要补充。
