Quantitative imaging of single mRNA splice variants in living cells

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

Research output: Contribution to journalArticlepeer-review

148 Scopus citations

Abstract

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.

Original languageEnglish
Pages (from-to)474-480
Number of pages7
JournalNature Nanotechnology
Volume9
Issue number6
DOIs
StatePublished - Jun 2014

Bibliographical note

Funding Information:
The authors thank Z. Machaty and K. Lee for the initial discussion on microinjection. The authors acknowledge funding to J.I. from the National Science Foundation (award no. 1249315), the Indiana Clinical Translational Sciences Institute (NIH-TR000006) and the Purdue Center for Cancer Research (NIH-NCI CCSG CA23168) and funding to K.L. and L.L. from Samsung (GRO 20122166) and Stanford NIH (award no. U54CA151459).

Fingerprint

Dive into the research topics of 'Quantitative imaging of single mRNA splice variants in living cells'. Together they form a unique fingerprint.

Cite this