Quantitative imaging of single mRNA splice variants in living cells.

Kyuwan Lee, Yi Cui, Luke P Lee, Joseph Irudayaraj

Nature nanotechnology 2014 Jun

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Alternative messenger RNA (mRNA) splicing is a fundamental process of gene regulation, and errors in RNA splicing are known to be associated with a variety of different diseases. However, there is currently a lack of quantitative technologies for monitoring mRNA splice variants in cells. Here, we show that a combination of plasmonic dimer probes and hyperspectral imaging can be used to detect and quantify mRNA splice variants in living cells. The probes are made from gold nanoparticles functionalized with oligonucleotides and can hybridize to specific mRNA sequences, forming nanoparticle dimers that exhibit distinct spectral shifts due to plasmonic coupling. With this approach, we show that the spatial and temporal distribution of three selected splice variants of the breast cancer susceptibility gene, BRCA1, can be monitored at single-copy resolution by measuring the hybridization dynamics of the nanoplasmonic dimers. Our study provides insights into RNA and its transport in living cells, which could improve our understanding of cellular protein complexes, pharmacogenomics, genetic diagnosis and gene therapies.

Citation

Kyuwan Lee, Yi Cui, Luke P Lee, Joseph Irudayaraj. Quantitative imaging of single mRNA splice variants in living cells. Nature nanotechnology. 2014 Jun;9(6):474-80