Synthetic-Molecule/Protein Hybrid Probe with Fluorogenic Switch for Live-Cell Imaging of DNA Methylation
Synthetic-Molecule/Protein Hybrid Probe with Fluorogenic Switch for Live-Cell Imaging of DNA Methylation
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DOI：
10.1021/jacs.7b09713
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发表时间：
2018-02-07
影响因子：
15
通讯作者：
Kikuchi, Kazuya
中科院分区：
文献类型：
--
作者：
Hori, Yuichiro;Otomura, Norimichi;Kikuchi, Kazuya
由合成分子和蛋白质组成的混合探针是检测活细胞中生物分子和信号的有力工具。迄今为止，大多数混合探针的目标仅限于pH值和小分析物。虽然生物大分子对细胞的生理功能至关重要，但基于混合探针的方法很少用于生物大分子的活细胞检测。在这里，我们开发了一种带有化学开关的杂交探针，用于甲基化DNA的活细胞成像，甲基化DNA是抑制基因表达的重要大分子。利用蛋白质标记技术，我们创建了一种混合探针，其中包含dna结合氟化物和甲基化dna结合结构域。杂交探针在与甲基化DNA结合时增强了荧光强度，并成功地监测了有丝分裂过程中的甲基化DNA。该杂交探针具有小分子或荧光蛋白探针所没有的显著优势，可用于与DNA甲基化相关的表观遗传现象和疾病的活细胞分析。
Hybrid probes consisting of synthetic molecules and proteins are powerful tools for detecting biological molecules and signals in living cells. To date, most targets of the hybrid probes have been limited to pH and small analytes. Although biomacromolecules are essential to the physiological function of cells, the hybrid-probe-based approach has been scarcely employed for live-cell detection of biomacromolecules. Here, we developed a hybrid probe with a chemical switch for live-cell imaging of methylated DNA, an important macromolecule in the repression of gene expression. Using a protein labeling technique, we created a hybrid probe containing a DNA-binding fluorogen and a methylated-DNA-binding domain. The hybrid probe enhanced fluorescence intensity upon binding to methylated DNA and successfully monitored methylated DNA during mitosis. The hybrid probe offers notable advantages absent from probes based on small molecules or fluorescent proteins and is useful for live-cell analyses of epigenetic phenomena and diseases related to DNA methylation.