模块化荧光互补传感器，用于活细胞检测内源基因组位点的表观遗传信号。,Nature Communications

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模块化荧光互补传感器，用于活细胞检测内源基因组位点的表观遗传信号。
Nature Communications ( IF 14.7 ) Pub Date : 2017-09-21 , DOI: 10.1038/s41467-017-00457-z
Cristiana Lungu ₁ , Sabine Pinter ₁ , Julian Broche ₁ , Philipp Rathert ₁ , Albert Jeltsch ₁

Affiliation

研究表观遗传过程的基本作用需要对活细胞中表观遗传修饰进行位点特异性检测的方法。在这里，我们通过开发四种用于活细胞显微镜应用的基于模块化荧光互补的表观遗传生物传感器来满足这一迫切需求。这些工具将工程化的 DNA 结合蛋白与识别定义的表观遗传标记的域结合起来，两者都与荧光蛋白的非荧光片段融合。目标 DNA 序列上表观遗传标记的存在导致功能性荧光团的重建。通过这种方法，我们首次能够直接检测活细胞内源基因组位点的 DNA 甲基化和组蛋白 3 赖氨酸 9 三甲基化，并跟踪这些标记在药物治疗、表观遗传酶诱导和细胞周期过程中的动态变化。我们预计这种多功能技术将提高我们对胚胎发育或疾病发作期间特定表观遗传特征如何设置、消除和维持的理解。表观遗传修饰成像工具可以揭示表观遗传过程的调节。在这里，作者提出了一种荧光互补方法，用于检测内源基因组位点的 DNA 和组蛋白甲基化，从而可以通过活细胞显微镜跟踪这些标记的动态变化。

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Modular fluorescence complementation sensors for live cell detection of epigenetic signals at endogenous genomic sites.

Investigation of the fundamental role of epigenetic processes requires methods for the locus-specific detection of epigenetic modifications in living cells. Here, we address this urgent demand by developing four modular fluorescence complementation-based epigenetic biosensors for live-cell microscopy applications. These tools combine engineered DNA-binding proteins with domains recognizing defined epigenetic marks, both fused to non-fluorescent fragments of a fluorescent protein. The presence of the epigenetic mark at the target DNA sequence leads to the reconstitution of a functional fluorophore. With this approach, we could for the first time directly detect DNA methylation and histone 3 lysine 9 trimethylation at endogenous genomic sites in live cells and follow dynamic changes in these marks upon drug treatment, induction of epigenetic enzymes and during the cell cycle. We anticipate that this versatile technology will improve our understanding of how specific epigenetic signatures are set, erased and maintained during embryonic development or disease onset.Tools for imaging epigenetic modifications can shed light on the regulation of epigenetic processes. Here, the authors present a fluorescence complementation approach for detection of DNA and histone methylation at endogenous genomic sites allowing following of dynamic changes of these marks by live-cell microscopy.

更新日期：2017-09-21

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