Development of direct, quantitative, and base-resolution sequencing methods for DNA and RNAmodifications

Thesis

DNA and RNA are decorated with various chemical modifications. Sequencing these modifications is crucial for us to understand their biological functions. To meet the growing demand for detecting DNA and RNA modifications, it is highly desirable to develop ideal chemical tools and sequencing methods that enable direct, quantitative, and base-resolution detection. In this thesis, I introduce the development of two such methods: Chemical-Assisted Pyridine borane Sequencing plus (CAPS+) for DNA 5-hydroxyethylcytosine (5hmC) sequencing and 2-Bromoacrylamide Assisted Cyclization Sequencing (BACS) for RNA pseudouridine (Ψ) sequencing. CAPS+ enhances the toolbox for sequencing DNA cytosine modifications, while BACS revolutionizes the Ψ detection. BACS has been applied to a series of studies on Ψ and pseudouridine synthases (PUS), revealing their functional importance in various biological processes and significantly deepening our understanding of this long known but enigmatic RNA modification. Overall, the development of CAPS+ and BACS clearly demonstrates the power of chemical tools in biological research. Further advancements in chemistry will continue to facilitate the study of DNA and RNA modifications.

Authors: Xu, H

• DOI: 10.5287/ora-kz966dwzb
• Type of award: DPhil
• Level of award: Doctoral
• Awarding institution: University of Oxford
• Language: English