Research scientist Donald Danforth Plant Science Center Chesterfield, Missouri
Body of Abstract: The sorting of RNA transcripts dictates their ultimate post-transcriptional fates, such as translation, decay, or degradation by RNA interference (RNAi). This sorting of RNAs into distinct fates is mediated by their interaction with RNA-binding proteins. Previously, we generated a protein-RNA tethering system that functions to drive an endogenous Arabidopsis RNA specifically into translation. Unlike other protein-RNA tethering systems that have been attempted in plants (dCas13, MS2, etc…), our system circumvents the inadvertent triggering of RNAi. We successfully tethered the translation factor to the endogenous target RNA, which functions to boost protein production in planta. To test if we can use this protein-RNA tethering system to investigate other RNA, we designed transgenic Cas9 RNA with different 3’UTRs for tethering, and in vivo tethered a translation-enhancing factor to a transgenic Cas9 RNA. Preliminary data shows that we can use this tethering system to increase Cas9 protein production and Cas9-mediated mutation rate by moving the translation-enhancing factor binding site to the Cas9 RNA. We have post-transcriptional engineered a protein-RNA tethering system and demonstrated that it is sufficient to direct the downstream fate of an exogenous RNA and boost protein production.