A Comprehensive Interrogation of Mirror-Image L-RNA-Protein Interactome Reveals Key Mechanisms of L-RNA Cytotoxicity

ABSTRACT: L-Oligonucleotides (ONs), the synthetic enantiomer of native D-nucleic acids, are being increasingly utilized for the development of diverse biomedical technologies, including molecular imaging tools, diagnostic biosensors, and aptamer-based therapeutics. Nevertheless, our understanding of how L-ONs behave in living systems falls far short of native D-ONs. In particular, despite the potential for an abundant L-ON-protein interactome, the extent to which L-ONs bind to endogenous proteins and the consequences of these interactions are unknown, posing a major hurdle towards engineering functional L-ONs with predictable intracellular behaviours. Towards closing this knowledge gap, we now report the first comprehensive characterization of L-ON–protein interactome, revealing that a wide-range nuclear proteins have the potential to bind L-RNA, especially those containing the common RNA recognition motif (RRM). Importantly, by focusing our study on cytotoxic L-RNA sequences, our data reveal key L-RNA–protein interactions that contribute to L-RNA cytotoxicity. Furthermore, we show that introducing 2-O-methyl modifications into cytotoxic L-RNA can decrease its cytotoxicity through reducing L-RNA–protein interactions, thereby demonstrating that a well-established strategy for mitigating the cytotoxic effects of antisense ONs (ASOs) can be translated across the chiral mirror. Overall, these findings greatly deepen our understanding of the intracellular behavior of L-ONs and provide valuable guidance for the future development of safe and effective L-ON-based biomedical technologies.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Epithelial Cell

DISEASE(S): Cervix Carcinoma

SUBMITTER: Xiaomei He  

LAB HEAD: Yinsheng Wang

PROVIDER: PXD054780 | Pride | 2025-03-13

REPOSITORIES: Pride