Click-encoded rolling FISH for visualizing single-cell RNA polyadenylation and structures.
IF: 19.160
Cited by: 13


Spatially resolved visualization of RNA processing and structures is important for better studying single-cell RNA function and landscape. However, currently available RNA imaging methods are limited to sequence analysis, and not capable of identifying RNA processing events and structures. Here, we developed click-encoded rolling FISH (ClickerFISH) for visualizing RNA polyadenylation and structures in single cells. In ClickerFISH, RNA 3' polyadenylation tails, single-stranded and duplex regions are chemically labeled with different clickable DNA barcodes. These barcodes then initiate DNA rolling amplification, generating repetitive templates for FISH to image their subcellular distributions. Combined with single-molecule FISH, the proposed strategy can also obtain quantitative information of RNA of interest. Finally, we found that RNA poly(A) tailing and higher-order structures are spatially organized in a cell type-specific style with cell-to-cell heterogeneity. We also explored their spatiotemporal patterns during cell cycle stages, and revealed the highly dynamic organization especially in S phase. This method will help clarify the spatiotemporal architecture of RNA polyadenylation and structures.


Gene Expression

MeSH terms

Cell Line, Tumor
Click Chemistry
DNA Barcoding, Taxonomic
Gene Expression Profiling
In Situ Hybridization, Fluorescence
MCF-7 Cells
Mass Spectrometry
Poly A
RNA, Messenger
Sequence Analysis, RNA
Single-Cell Analysis
Spatio-Temporal Analysis


Chen, Feng
Bai, Min
Cao, Xiaowen
Zhao, Yue
Xue, Jing
Zhao, Yongxi

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