Danio rerio Raw sequence reads. Danio rerio

To investigate the roles of p15PAF in zebrafish development, we generated p15PAF mutant zebrafish using CRISPR/Cas9 technology. Maternal loss of zebrafish p15PAF (p15PAF-M) led to severe defects in early embryonic development including axis formation that was indicated by reduced or dislocated expression of a number of marker genes for early embryonic dorsoventral patterning. Strikingly, the normal degradation of maternal factors and activation of zygotic genes during maternal-to-zygotic transition (MZT) were severely disturbed, such as the down-regulated expression of several components of the Wnt/β-catenin signaling pathway. Interestingly, the dorsal-ventral patterning defects in p15PAF-M embryos were rescued by ectopic expression of β-catenin. The TOPFlash luciferase reporter assay that was used to analyze TCF/LEF activity of Wnt/β-catenin signaling showed that p15PAF and β-catenin cooperate to regulate the downstream genes. Co-immunoprecipitation assays confirmed that zebrafish P15PAF could bind to β-catenin. In summary, our results reveal that zebrafish p15PAF interacts with β-catenin and regulates its activity and plays important roles in early embryonic development including MZT and axis formation.