Research Interests
Our lab aims to develop RNA-based tools to manipulate pathways within living cells. We design synthetic RNA molecules – including RNA aptamers and circular RNAs – to serve as precise tools for interrogating and manipulating cellular processes. By integrating these RNA-based tools with approaches from biochemistry, molecular biology, and cell biology, we seek to answer fundamental biological questions and pioneer novel strategies for RNA-based therapeutics.
1.Programming Cellular Pathways with Synthetic RNA
Intermolecular interactions play crucial roles in many cellular activities, including enzymatic reactions, signal transduction, and gene expression. However, precise control of these interactions remains a significant challenge. Our previous work demonstrated that modular RNA molecules (like dual-specificity aptamers) can be deployed to regulate glycosylation of a designated protein by inducing protein-protein interactions (Zhu and Hart, 2023). We aim to expand this platform to regulate diverse cellular pathways and to discover mechanisms that control these molecules with high efficacy and specificity. Our research will provide new tools for basic science, and lay the foundation for a novel class of RNA-based therapeutics.
2.Dissecting the O-GlcNAc Regulation in Health and Disease
O-GlcNAc (O-linked b-N-acetylglucosamine) is a post-translational modification that regulates protein functions. It involves a single GlcNAc molecule attached to the hydroxyl group of a serine (Ser) or threonine (Thr) residue. O-GlcNAc has been identified on more than 9000 proteins in human cells. It regulates various protein functions, including enzymatic activity, protein-protein interactions, and protein degradation. However, our knowledge of how O-GlcNAc regulates individual protein function remains limited. Using RNA tools and other approaches, we aim to dissect how this abundant sugar modification controls specific protein functions, and how abnormal O-GlcNAcylation contributes to human diseases such as cancer, diabetes, and neurodegenerative diseases.
3.Innovating Circular RNA Technology
Circular RNAs (circRNAs) are single-stranded RNA molecules whose 5’ and 3’ ends are covalently joined. Without free ends, circRNAs are naturally resistant to exonuclease degradation and are exceptionally stable. These properties make circRNA a promising platform for the design of RNA tools, and for the durable expression of therapeutic proteins and vaccines in vivo. However, the full potential of circRNA is hindered by inefficient production, cumbersome purification, and limited protein translation efficiency. We aim to overcome these limitations by developing novel production and purification strategies, and by optimizing circRNA sequences to enhance protein expression. Our goal is to create a robust, efficient, and user-friendly circRNA platform to significantly expand its research and therapeutic applications.
Publications
1.Zhu, Y.*, and Hart, G.W.* (2023). Dual-specificity RNA aptamers enable manipulation of target-specific O-GlcNAcylation and unveil functions of O-GlcNAc on b-catenin. Cell. doi:10.1016/j.cell.2022.12.016. Co-corresponding Author.
Highlighted by:
Cheng, S.S., and Woo, C.M. (2023). Dual-specificity RNA aptamers: A sweet new tool for studying O-GlcNAc biology. Mol Cell. doi:10.1016/j.molcel.2023.01.025.
Pratt, M.R. (2023). A sticky solution to protein-selective sugar installation. Cell Res. doi:10.1038/s41422-023-00787-2.
2.Zhu, Y., and Hart, G.W. (2021). Nutrient regulation of the flow of genetic information by O-GlcNAcylation. Biochem Soc Trans. doi:10.1042/BST20200769.
3.Zhu, Y., and Hart, G.W. (2020). Targeting O-GlcNAcylation to develop novel therapeutics. Mol Aspects Med. doi:10.1016/j.mam.2020.100885.
Patents
1.Gerald W. Hart; Yi Zhu; DUAL-SPECIFICITY RNA APTAMERS FOR REGULATING O-GLCNACYLATION, 2023-05-25, WIPO, PCT/US2022/079729；2024-04-16, United States, 18/701,725
