Date of Award

12-2025

Degree Type

Dissertation

Degree Name

Doctor of Philosophy (PhD)

School

Mathematics and Natural Sciences

Committee Chair

Faqing Huang

Committee Chair School

Mathematics and Natural Sciences

Committee Member 2

Vijay Rangachari

Committee Member 2 School

Mathematics and Natural Sciences

Committee Member 3

Yanlin Guo

Committee Member 3 School

Biological, Environmental, and Earth Sciences

Committee Member 4

Alex Flynt

Committee Member 5

Jacques Kessl

Committee Member 5 School

Mathematics and Natural Sciences

Abstract

Though once considered as simply an intermediate form of gene expression, the myriad physiological roles of RNA elucidated over the last several decades have wholly transformed the scientific community’s understanding of RNA biochemistry and its significance in disease and therapeutics. As RNA modifications have been demonstrated to exert a plethora of roles integral to RNA function and fate, further illumination of RNA modifications, their biogenesis mechanisms, their physiological purposes, their therapeutic utilities, and their evolutionary descent have approached the forefront of modern RNA research. While the m7G cap was long believed to be the sole RNA cap and specific to eukaryotes, groundbreaking 2009 studies provided the first evidence for the previously-hypothesized existence of noncanonical caps such as NAD and CoA. As reliable methods for isolating intact NAD-capped RNAs swiftly followed these initial reports, much has been elucidated regarding NAD-RNA such as its biogenesis mechanisms, physiological titers, and physiological roles in both prokaryotes and eukaryotes. However, the biological synthesis and purposes of FAD-capped RNAs have largely remained undetermined, with less than a handful of reports to date describing FAD-RNA detection and only a single report detailing physiological roles and synthesis. This deficiency in FAD-RNA investigations is largely due to the lack of reliable methods for isolating intact FAD-RNAs. Here, we report a flavin analog-based method for the isolation of intact FAD-capped RNAs, as well as strategies and preparations for utilizing this method to identify FAD-RNAs generated in vivo and perform SELEX for FMN-specific FAD synthetase ribozymes.

Available for download on Thursday, December 31, 2026

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