Date of Award

2024

Document Type

Open Access Dissertation

Degree Name

Doctor of Philosophy in Biochemistry and Molecular Biology (PhD)

Administrative Home Department

Department of Chemistry

Advisor 1

Shiyue Fang

Advisor 2

Yinan Yuan

Committee Member 1

Marina Tanasova

Committee Member 2

Xiaohu (Mark) Tang

Committee Member 3

Tarun Dam

Abstract

In the field of epigenetics - the dynamic collection of chemical modifications that overlay DNA/RNA - there is increasing interest in the potential for therapeutic and research-based utilization of the epitranscriptomics: those epigenetic modifications specific to RNA. Epitranscriptomics can give us a new approach to understanding and treating RNA viruses, like SARS-CoV-2. Given viruses are often found modified by human epitranscriptomic machinery, understanding how these modifications can modulate virulence is of high interest. We have shown that the N1-methyladensoine modification causes RNA synthesis termination in the SARS-CoV-2 replication complex, and thus may explain the differences in symptoms observed in individuals. Furthermore, we proved that this modification can be incorporated into RNA via its triphosphate form using the same replication complex, and thus could serve as an effective natural antiviral therapeutic to SARS-CoV-2 and other coronaviruses. Sensitive epitranscriptomic modifications (e.g., ac4C) are yet another category of epigenetics which are similarly of high interest to biological research. However, the chemical synthesis of RNA containing sensitive epitranscriptomic modifications is extremely limited. The standard synthesis, cleavage and deprotection, methods currently used cannot synthesize RNA with sensitive groups. This is chiefly due to the harsh basic conditions needed to deprotect and cleave the RNA, following synthesis. Attempts at creating new protecting groups and solid support linkers have made some progress in the synthesis of sensitive RNA, but their methods cannot produce RNA of acceptable length or purity due to lowered coupling yields and cumbersome photocleavage steps. We have developed a novel silyl containing cyclizable protecting group (SoM) and linker (SoA) that have achieved RNA synthesis at the length of 26 nucleotides that retained the sensitive modification ac4C in good purity and yields.

Creative Commons License

Creative Commons Attribution 4.0 License
This work is licensed under a Creative Commons Attribution 4.0 License.

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