(644.7) Defining the role of RNA acetylation via a sequence-specific RNA acetyltransferase

Poster Board Number: A34

Samantha Nance (National Cancer Institute), Jordan Meier (National Cancer Institute), Raj Chari (National Cancer Institute), Christine Evans (National Cancer Institute)

RNA modifications play a critical role in protein abundance, biology, and disease. However, methods for testing the causal effects of individual RNA modifications remain in their infancy. Here we report the preliminary development of an epitranscriptome engineering strategy to manipulate RNA cytidine acetylation in living cells. Our approach fuses a eukaryotic cytidine acetyltransferase to a sequence-specific RNA-binding protein. First, we optimize the ectopic expression of these constructs in human embryonic kidney cells. Next, we develop and apply assays to assess guide-dependent and independent acetylation of a model substrate. Finally, we demonstrate the dependence of modification on consensus sequence, guide/target proximity, and linker protein fusion geometry. These studies provide a foundation for site-specific rescue of individual cytidine acetylation events in model organisms, and in so doing will help illuminate the function of RNA modifications in disease.