Dissecting the mechanisms of miRNA function in establishment of neuronal polarity

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Neuron is a highly polarized cell formed by a cell body, or soma, and cell extensions, or neurites. Such polarity is crucial to proper neuronal function. Establishment of cell polarity in large relies on asymmetric subcellular localization and translation of mRNAs. In our prior work, we have developed neurite/soma fractionation scheme in combination with mass spectrometry, RNA-seq, Ribo-seq and bioinformatics analyses, to identify proteins and RNAs that are differentially localized and translated between neurites and soma of neuronal cells. Moreover, we showed that mRNA localization and local translation are strongly affected by abrogation of the microRNA (miRNA) pathway. miRNAs are important regulators of gene expression with known roles in translational repression and mRNA decay, targeting more than a half of all eukaryotic genes. However their functions in subcellular localization and translation of mRNAs have not been systematically analyzed. In the proposed study, we aim to understand the network of miRNA-mediated regulation of mRNA localization and translation patterns in neuronal cells. We will map miRNA-binding sites in localized and locally translated mRNAs using AGO PAR-CLIP, perform integrative computational data analysis to identify miRNAs that regulate subcellular localization and translation patterns and dissect the underlying mechanisms of action. The proposed research has the potential to provide important insights into the mechanisms underlying establishment of cell polarity and roles of miRNAs in this process.