Deficiency of acetyltransferase nat10 in zebrafish causes developmental defects in the visual function and behavior

N4-acetylcytidine (ac4C) is an epitranscriptomic modification of mRNA that is catalyzed by N-acetyltransferase 10 (NAT10), a critical factor known to influence mRNA stability. However, its role in development has not been investigated. In this study, we used CRISPR/Cas9 and RNAi technology to knock out and knock down nat10, the zebrafish ortholog of human NAT10, and evaluated their effects on development, behavior, and transcriptome. Our findings indicate that nat10 deficiency in zebrafish embryos results in increased embryo mortality and developmental abnormalities. Additionally, behavioral and histological evaluations revealed that nat10 knockdown led to increased anxiety-like behavior and severe vision impairment. Transcriptome profiling and RT-PCR results showed that nat10 knockdown significantly downregulated the expression of retinal transcripts that are enriched in response to light stimuli, photoreceptors, and visual perception. Furthermore, dot-blot and RIP-PCR analyses confirmed a significant reduction in ac4C levels in total RNA and opsin mRNA in nat10 knockdown zebrafish. Our results highlight the essential role of ac4C in embryonic development, especially in visual development. This zebrafish model could be helpful for studying ac4C modification in neurodevelopmental disorders.