Detection of RNA-DNA association by a proximity ligation-based method.
Detection of RNA-DNA association by a proximity ligation-based method.
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通过基于邻近连接的方法检测 RNA-DNA 关联。
DOI：
10.1038/srep27313
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发表时间：
2016
影响因子：
4.6
通讯作者：
Mazo,Alexander
中科院分区：
文献类型：
--
作者：
Petruk,Svetlana;Fenstermaker,TylerK;Black,KathrynL;Brock,HughW;Mazo,Alexander
我们描述了一种基于邻近连接测定 (PLA) 的方法，用于评估细胞周期中单细胞中 DNA 和 RNA 的关联。用 EdU 对 DNA 进行脉冲标记，用 BrU 对 RNA 进行脉冲标记，并用 PLA 测试它们的接近程度，结果表明单个细胞中的 RNA 合成在 DNA 复制后约 30-45 分钟恢复。与这一结论一致的是，在其 CTD 的 Ser2 处磷酸化的 RNA Pol II 与新生 DNA 上的 RNA 转录物同时被检测到。我们的结果还表明，RNA 在有丝分裂的所有阶段都与 DNA 焦点相关。
We describe a proximity ligation assay (PLA)-based method of assessing association of DNA and RNA in single cells during the cell cycle. Pulse-labeling of DNA with EdU and RNA with BrU and testing their close proximity by PLA demonstrates that RNA synthesis in individual cells resumes about 30–45 min after DNA replication. Consistent with this conclusion, RNA Pol II phosphorylated at Ser2 of its CTD is detected at the same time as RNA transcripts on nascent DNA. Our results also show that RNA is associated with DNA foci during all stages of mitosis.