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Development of cell-specific RNA metabolic labeling applications in cell and animal models


The development of new tools for RNA metabolic labeling is critical for analyzing nascent transcription in cells and animal models to elucidate specific regulatory mechanisms of RNA. These approaches have recently been demonstrated with commercially available methods, such as thiol (SH)-linked alkylation for the metabolic sequencing of RNA (SLAM-seq), yet there remain many opportunities to refine RNA metabolic labeling for cell-specific approaches. In the following dissertation, I will highlight the important work that has already been developed towards these efforts. I will also introduce a novel cell-specific RNA metabolic labeling method using a new enzyme-analog pair. For this approach, I will demonstrate how it can be used in vivo to capture nascent RNA within a metastatic breast cancer mouse model. Lastly, I will discuss a new chemo-selective reagent pair for vinyl nucleosides that can be used to analyze two RNA populations simultaneously in cells. Throughout the following document, I will emphasize the chemical and biochemical challenges and advantages of these labeling methods and discuss opportunities for future development. These tools are dedicated to improving biological insight to study transcription with greater physiological specificity.

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