Due to the great complexity of mRNA translation and the diversity in mRNA populations from one cell to another, tools for studying gene expression at the single-cell level are increasingly in demand.  This is particularly true in analysis of complex tissues where cell-cell interactions and cellular position elaborate functional pathways.  To overcome this issue and to isolate RNA from single cells in their natural environment, a novel photochemical approach for isolating mRNA from single neurons was developed.  This approach has been called Transcriptome In Vivo Analysis or TIVA-tag.  This design utilizes synthesis of a photoactivatable oligonucleotide hairpin with an antisense strand that targets the poly-A tails of mRNA.  When introduced into cells and activated, the TIVA-tag compound binds to endogenous poly-A+ RNA permitting its isolation.  Experiments in single neurons have confirmed the utility of the hairpin for the isolation of mRNA from single cells.

Transcriptome in vivo analysis: A photoregulatable approach to mRNA isolation

243rd National Meeting (2012)

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