EJC impact on mRNA localization and packaging – Eat_local

mRNA intracellular localization is widespread. It favors local expression of specific proteins contributing to cell polarity and differentiation. Recently, we described the first vertebrate mRNA whose localization depends on the exon-junction complex (EJC). Loaded onto spliced exon junctions, EJCs are key for the mRNA lifecycle and our data suggest a transcript-specific EJC pattern. Using human and Drosophila edited cell lines and cutting-edge methods, we will address the role of EJC in mRNP packaging and localization, by: 1) determining the EJC deposition code using RNA cross-linking methods with improved sensitivity; 2) screening for new EJC-dependent localized transcripts by smFISH and proximity assays; 3) characterizing cis- and trans-acting signals involving EJC in mRNA localization, in relation to translation; 4) examining the role of EJC on RNA conformation, both in vitro by single nucleic acid molecule manipulation and in vivo by smFISH.