Fusarium infection alters the m⁶A-modified transcript landscape in the cornea.

N6-methyladenosine (m⁶A) is the most common post-transcriptional modification of RNA in eukaryotes that regulates the post-transcriptional expression level of genes without changing the base sequence. The role of m⁶A in fungal keratitis has not yet been elucidated. Here, we aimed to identify m⁶A modification changes and their potential roles in fungal keratitis. The murine model of fungal keratitis was established by inoculating mice with Fusarium solani (F. solani). The overall m⁶A level was detected via an m⁶A RNA methylation assay kit. The expression levels of key m⁶A modification-related genes were estimated by quantitative real-time polymerase chain reaction (PCR). The expression and localization of METTL (methyltransferase like)3, the key component of the m⁶A methyltransferase complex, was determined by immunostaining and Western blotting (WB). Immunoprecipitation methylation microarray was used to describe the changes in m⁶A modification in F. solani-infected corneal tissue. The overall m⁶A level in corneal tissue on the 5th day in the F. solani-treated group was upregulated compared with that in the control group. The demethylase levels were unaltered, but the level of the methylase METTL3 was increased significantly after fungal infection. Additionally, differences were found in m⁶A modifications in 1137 mRNAs, of which 780 were hypermethylated and 357 were hypomethylated. To the best of our knowledge, the present work is the first investigation on the m⁶A modification profiles in experimental fungal keratitis, and it may provide a potential therapeutic target.

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