Oral Presentation Australasian RNA Biology and Biotechnology Association 2026 Conference

RNA methylation, particularly N6-methyladenosine (m⁶A), plays a critical role in diverse physiological processes and human diseases. For decades, our understanding of m⁶A function has largely relied on perturbing the proteins responsible for depositing, recognising, or erasing this modification. While these approaches have provided important biological insights, they induce global alterations to the epitranscriptome and can be confounded by effects on cell viability, transcriptome-wide changes, and other off-target phenotypes. The development of CRISPR-Cas-based RNA m⁶A editing technologies has created new opportunities to directly investigate the functional consequences of site-specific m⁶A modifications. By enabling precise manipulation of individual methylation sites, these tools offer a powerful framework for establishing causal links between m⁶A deposition and biological outcomes. However, programmable m⁶A editing experiments are associated with several technical and conceptual challenges. Without rigorous experimental design and appropriate controls, such studies may generate misleading or spurious conclusions regarding the role of specific m⁶A sites. This presentation will discuss the importance of programmable epitranscriptome editing, highlight common pitfalls in the design and interpretation of m⁶A editing experiments, and outline critical experimental considerations and best-practice strategies to ensure robust, reproducible, and biologically meaningful conclusions.