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Issue 5, 2017
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Highly specific imaging of mRNA in single cells by target RNA-initiated rolling circle amplification

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Abstract

Detection of single-cell gene expression with high spatial and sequence resolution is a key challenge in single cell biology. Herein, we propose a robust method for the direct detection of mRNA, termed target RNA-initiated rolling circle amplification, which enables imaging of mRNA with single-nucleotide and near-single-molecule resolution in single cells. By utilizing a Splint R ligase capable of efficiently catalyzing the ligation of a padlock probe by the target RNA, the method can enable the efficient detection of mRNA without reverse transcription (detection efficiency over 20%). Meanwhile, attributed to the ligation-based recognition process, the method confers specificity sufficient to genotype mRNAs with one-nucleotide variations. The method has enabled the spatial mapping and correlation analysis of gene expression in single cells which could help us to elucidate the gene functions and regulatory mechanisms. This method offers an mRNA profiling ability with high spatial resolution and sequence specificity, thus is expected to be a single-cell analysis platform for not only investigating gene expression, but also potentially for analyzing single-nucleotide variants or mRNA alternative splicing at single-cell level.

Graphical abstract: Highly specific imaging of mRNA in single cells by target RNA-initiated rolling circle amplification

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Publication details

The article was received on 20 Jan 2017, accepted on 05 Mar 2017 and first published on 07 Mar 2017


Article type: Edge Article
DOI: 10.1039/C7SC00292K
Citation: Chem. Sci., 2017,8, 3668-3675
  • Open access: Creative Commons BY license
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    Highly specific imaging of mRNA in single cells by target RNA-initiated rolling circle amplification

    R. Deng, K. Zhang, Y. Sun, X. Ren and J. Li, Chem. Sci., 2017, 8, 3668
    DOI: 10.1039/C7SC00292K

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