Institute of Molecular Medicine , Peking University , 100871 Beijing , China.
Research Institute of Dalian University of Technology in Shenzhen , Gaoxin South fourth Road , Nanshan District, Shenzhen 518057 , China.
Anal Chem. 2019 Mar 5;91(5):3336-3341. doi: 10.1021/acs.analchem.8b04418. Epub 2019 Feb 14.
Many mitochondria-related diseases are associated with the mutation of mitochondrial DNA (mtDNA). Therefore, visualizing its dynamics in live cells is essential for the understanding of the function of mtDNA transcription and translation. By employing carbazole as the framework and designing a module for DNA minor-groove binding, here we have developed a novel fluorescent probe with a large Stokes shift (λ = 480 nm and λ = 620 nm), CNQ, for mtDNA detection and visualization. It is almost nonfluorescent in PBS buffer and exhibits 182-fold enhancement in fluorescence within 20 s after the application of mtDNA in the solution, with a detection limit of 55.1 μg/L. Using dual-color Hessian-structured illumination microscopy, we have demonstrated that CNQ-labeled mtDNA structures are distinct from those labeled by TFAM-EGFP. Finally, we have used two-photon confocal scanning microscopy (λ = 850 nm) to monitor the nondestructive doxorubicin-induced mtDNA damage in live cells.
许多与线粒体相关的疾病都与线粒体 DNA（mtDNA）的突变有关。因此，在活细胞中可视化其动态对于理解 mtDNA 转录和翻译的功能至关重要。通过以咔唑为骨架，并设计一个用于 DNA 小沟结合的模块，我们开发了一种新型荧光探针 CNQ，用于 mtDNA 的检测和可视化。在 PBS 缓冲液中，它几乎没有荧光，在溶液中加入 mtDNA 后的 20 秒内，其荧光强度增强了 182 倍，检测限为 55.1μg/L。使用双色彩虹结构照明显微镜，我们证明了 CNQ 标记的 mtDNA 结构与 TFAM-EGFP 标记的 mtDNA 结构不同。最后，我们使用双光子共聚焦扫描显微镜（λ=850nm）监测活细胞中阿霉素诱导的 mtDNA 损伤的非破坏性。
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