Visualizing Newly Synthesized RNA by Bioorthogonal Labeling-Primed DNA Amplification
Visualizing Newly Synthesized RNA by Bioorthogonal Labeling-Primed DNA Amplification
复制标题
通过生物正交标记引发的 DNA 扩增可视化新合成的 RNA
DOI：
10.1021/acs.analchem.0c01161
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发表时间：
2020
影响因子：
7.4
通讯作者：
Zhao Yongxi
中科院分区：
文献类型：
--
作者：
Bai Min;Cao Xiaowen;Huang Wei;Chen Feng;Zhao Yue;Xue Jing;Li Youjun;Cheng Yilong;Zhang Lei;Zhao Yongxi
Monitoring RNA synthesis and spatial distribution can help to understand its role in physiology and diseases. However, visualizing newly synthesized RNA in single cells remains a great challenge. Here, we developed a bioorthogonal labeling-primed DNA amplification strategy to visualize newly synthesized RNA in single cells. The new bioorthogonalN6-allyladenosine nucleoside was prepared to metabolically label cellular newly synthesized RNAs. These allyl-functionalized RNAs then reacted with tetrazine-modified primers. These primers could initiate rolling circle amplification, producing tandem periodic long single DNA strands to capture hundreds of fluorescence probes for signal amplification. Using this method, we explored the subcellular distributions of newly synthesized RNAs. And we found that newly synthesized RNAs are spatially organized in a cell type-specific style with cell-to-cell heterogeneity.
