Characterization of prps1a gene editing and phenotypic analysis in zebrafish. (a) Schematic of the zebrafish prps1a gene structure, highlighting the gRNA target sites. Arrows indicate gRNA binding sites, with exons represented as boxes. (b) Sequencing analysis of mutations induced by the two gRNAs showing various mutation types, including insertions, deletions, and substitutions. (c) Validation of gRNA activity through cleavage assays, assessing the efficiency of double-strand break induction by each gRNA. (d) Hair cell staining in zebrafish larvae across four experimental groups: wild-type (WT), gene KO, KO with wild-type mRNA injection (KO+WT mRNA), and WT with Cys165Tyr mutant mRNA injection (WT+Cys165Tyr). Scale bar = 200 μm. (e) Confocal microscopy images of hair cells in zebrafish larvae from different experimental groups showing structural integrity and localization. Scale bar = 20 μm. (f) Quantification of hair cells in different experimental groups. (g) Behavioral tracking of zebrafish larvae, recording movement patterns to assess the impact of prps1a gene editing on behavior. (h) Line graph of swimming distance of zebrafish with different genotypes under noise. (i) Statistical analysis of behavioral data of total distance. (j) Statistical analysis of behavioral data of first distance. (k) Representative microscopic images of hair cells stained with AO in zebrafish larvae from different experimental groups. Scale bar = 100 μm. (l) Quantification of fluorescence intensity in hair cells of the larvae. Data were compared using ANOVA followed by post hoc tests, with significant differences indicated (p < 0.05). Data are presented as mean ± SD. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001.
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