Aberrant RNA localization plays a role in many diseases including cancer, Alzheimer's, and Huntington's disease. Therefore, developing techniques for visualizing mRNA localization in cells would provide insight into these diseases. Currently, the most popular method for labeling mRNA involves using a genetically encoded green fluorescent protein (GFP) tag. This allows for direct visualization of mRNA in the cell.However, the protein fusion is large and can disrupt normal mRNA transport in the cell and native mRNA structure.We have developed a ribozyme that covalently self-alkylates with fluorescein and could be fused to an mRNA sequence of interest. Benefits of this covalent linkage include providing a robust complex that prevents signal loss, the ability to wash out any unbound fluorescein and reduce background fluorescence, and a broader fluorophore scope. This method may enable the rapid screening of small molecules for efficacy in modulating cellular RNA localization.A general scheme of ribozyme labeling with a fluorescent probe, where x represents a leaving group.

Covalent mRNA labeling using fluorescently self-alkylating ribozymes

248th National Meeting (2014)

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