Protein-PNA conjugates as conditional BRET platforms for life cell mRNA imaging and quantification

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Within the framework of this proposal, a new highly sensitive imaging method will be developed to enable visualization and spatial quantification of low abundance mRNA isoforms in cells in real time. Despite the variety of existing RNA imaging methods, it is major hurdle to study mRNA isoforms in real time in living cells due to the lack of sensitivity of current strategies. Importantly, these low abundance transcripts formed mainly by alternative splicing contribute significantly to protein diversity and play a central role in development, neuronal plasticity, and nervous system disorders such as Alzheimer's, spinal muscular atrophy (SMA), etc. Based on the well-characterized mRNA isoforms responsible of SMA, we aim to develop a PNA-NanoBRET bioimaging platform, which allows follow gene expression in real time in living cells. This novel method will ultimately be tested in genetically modified cells and patient fibroblasts.