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The study of RNA function is central to contemporary biology as it plays important roles in cellular functions such as patterning, growth and differentiation in many organisms. In Drosophila, posterior localization of oskar RNA in the oocyte is necessary for germ line determination and abdomen formation in the embryo. Although splicing and the exon junction complex (EJC) proteins are genetically required for its transport, the mechanistic understanding of the role of nuclear events in assembly of the oskar RNP complex was lacking. Using in vivo transgenic approach, I identified a novel bipartite cis-acting element in the oskar coding region that is formed upon splicing and together with the EJC proteins, is necessary for posterior localization. Structure-function analysis supported the formation of a RNA structure whose integrity alone is critical for localization. Time lapse imaging of the in vivo tagged RNA highlighted the role of the cis-element in recruitment and/or stabilization of kinesin on oskar RNP for posterior transport.
Furthermore, biochemical analysis of the EJC-interacting protein PYM revealed its role in EJC homeostasis and RNA transport in the oocyte.
Finally, I will present data showing the use of CRISPR technology to study long non-coding RNA functions which can be adapted to program gene expression in vivo.
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