Analysis of RNAs in their endogenous environment could provide unique insights into cell-type-specific gene expression and RNA dynamics. However, visualizing individual RNA molecules within or isolating RNAs from a live single cell has remained challenging. Two recent reports look to drive the field forward in that respect. Park et al. engineered a transgenic mouse model that expresses fluorescently labelled endogenous β-actin mRNA to visualize its localization and movement. Experiments in cultured neurons and acute brain slices showed that β-actin mRNA molecules undergo continuous assembly and disassembly in large mRNA–protein complexes. The authors believe that their live-cell imaging technique is applicable to other genes to investigate the dynamic regulation of individual components of the transcriptome. In the second study, Lovatt et al. describe an mRNA capture methodology that enables the isolation of mRNA from live single cells in intact complex tissues. They engineered a chemical compound called the TIVA (transcriptome in vivo analysis) tag that enters live cells and, upon photoactivation, anneals to mRNA. TIVA–mRNA hybrids can be isolated through an affinity tag incorporated into the TIVA tag, and subsequent RNA sequencing thus yields the transcriptome profile of a single cell.
References
Park, H. Y. et al. Visualization of dynamics of single endogenous mRNA labeled in live mouse. Science 343, 422–424 (2014)
Lovatt, D. et al. Transcriptome in vivo analysis (TIVA) of spatially defined single cells in live tissue. Nature Methods 11, 190–196 (2014)
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Koch, L. Tagging mRNAs in living cells. Nat Rev Genet 15, 144 (2014). https://doi.org/10.1038/nrg3691
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DOI: https://doi.org/10.1038/nrg3691