Probing Transient DNA Conformation Changes with an Intercalative Fluorescent Excimer
Probing Transient DNA Conformation Changes with an Intercalative Fluorescent Excimer
复制标题
使用嵌入荧光准分子探测瞬时 DNA 构象变化
DOI：
10.1002/anie.202014466
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发表时间：
2021-02-03
影响因子：
16.6
通讯作者：
Li, Jiang
中科院分区：
文献类型：
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作者：
Chen, Bin;Huang, Qiuling;Li, Jiang
DNA构象的变化在调节基因表达和介导药物-DNA相互作用中是重要的。然而，由于该过程的动态性质，直接探测瞬时DNA构象变化具有挑战性。我们展示了一种无标记的荧光方法来监测具有不同长度和形状的DNA结构中的瞬时DNA构象变化，使用DNA嵌入剂K21。K21可以在DNA表面形成瞬时激基体，单体和激基体的比率发射与许多DNA结构中的DNA瞬时构象稳定性相关，包括i-基序、G-四链体结构和随机位置的单核苷酸突变。我们分析了一个单一的质粒酶切前后的构象动力学与共聚焦荧光显微镜。该方法提供了一种无标记的荧光策略来探测DNA结构的瞬时构象变化，并具有揭示活细胞中瞬时基因组过程的潜力。
Variation of DNA conformation is important in regulating gene expression and mediating drug-DNA interactions. However, directly probing transient DNA conformation changes is challenging owing to the dynamic nature of this process. We show a label-free fluorescence method to monitor transient DNA conformation changes in DNA structures with various lengths and shapes using a DNA intercalator, K21. K21 can form transient excimers on the surface of DNA; the ratiometric emission of monomer and excimer correlate to DNA transient conformation stability in numerous DNA structures, including i-motifs, G-quadruplex structures, and single nucleotide mutation at random position. We analyzed the conformation dynamics of a single plasmid before and after enzyme digestion with confocal fluorescence microscopy. This method provides a label-free fluorescence strategy to probe transient conformation changes of DNA structures and has potential in uncovering transient genomic processes in living cells.