Pairwise Proximity-differentiated Visualization of Single-cell DNA Epigenetic Marks
Pairwise Proximity-differentiated Visualization of Single-cell DNA Epigenetic Marks
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单细胞 DNA 表观遗传标记的成对邻近差异可视化
DOI：
10.1002/anie.202011172
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发表时间：
2020
期刊：
影响因子：
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通讯作者：
Zhao Yongxi
中科院分区：
文献类型：
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作者：
Xue Jing;Chen Feng;Su Li;Cao Xiaowen;Bai Min;Zhao Yue;Fan Chunhai;Zhao Yongxi
空间定位和相关生物分子的接近，如DNA表观遗传标记，是深入了解生命的基础。然而，它仍然很少探索，技术上具有挑战性。在这里，我们报告了单细胞5-甲酰胞嘧啶（5 fC）和5-羟甲基胞嘧啶（5 hmC）的成对邻近分化可视化。固定细胞染色质上的这两个标记通过点击化学连续标记并与它们的DNA引物探针交联。基于成对邻近分化机制，近端5 fC/5 hmC位点和残余5 fC或5 hmC位点用相应的环化条形码编码。这些条形码同时扩增用于多重单分子成像。因此，我们证明了5 fC或5 hmC空间定位和它们在单细胞中的成对接近的差异化可视化。这种多水平的亚细胞信息可以提供对染色质修饰的调节功能和机制的见解，并且空间接近性可以暴露其阅读器蛋白之间的潜在串扰或相互作用。
Spatial positioning and proximity of relevant biomolecules such as DNA epigenetic marks are fundamental to a deeper understanding of life. However, it remains poorly explored and technically challenging. Here we report the pairwise proximity‐differentiated visualization of single‐cell 5‐formylcytosine (5fC) and 5‐hydroxymethylcytosine (5hmC). These two marks on chromatin in fixed cells are successively labeled and crosslinked with their DNA primer probes via click chemistry. Based on a pairwise proximity‐differentiated mechanism, proximal 5fC/5hmC sites and residual 5fC or 5hmC sites are encoded with respective circularized barcodes. These barcodes are simultaneously amplified for multiplexed single‐molecule imaging. We thus demonstrate the differentiated visualization of 5fC or 5hmC spatial positioning and their pairwise proximity in single cells. Such multi‐level subcellular information may provide insights into regulation functions and mechanisms of chromatin modifications, and the spatial proximity can expose the potential crosstalk or interaction between their reader proteins.