Characterization of co-Regulatory RNA Splicing Targets of ALKBH5 and FTO

Abstract

Alternative splicing is a key gene regulatory mechanism in eukaryotic cells that generates diverse mRNA and protein isoforms. N6-methyladenosine (m6A), a reversible RNA modification mediated by writer complexes and eraser proteins, plays a critical role in regulating alternative splicing. Among the known eraser proteins, ALKBH5 and FTO are well-characterized m6A demethylases. Given that both proteins catalyze m6A demethylation, I hypothesized that there may be a subset of genes whose splicing is co-regulated by both ALKBH5 and FTO. To investigate this, I conducted a bioinformatic analysis and identified a set of candidate genes potentially co-regulated by these two demethylases. To validate these computational findings, I performed RT-PCR on representative candidate genes selected based on P-value significance and sashimi plot result. The RT-PCR result confirmed that alternative splicing changes occurred in these genes. Overall, this study serves as a preliminary investigation aimed at exploring how demethylases regulate RNA splicing. Its main significance lies in the identification and validation of candidate genes that are co- regulated by both ALKBH5 and FTO.