ZFIN Image: Trejo-Reveles et al., 2023, Figure 4

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Figure 4

Targeted gene disruption of OFC novel candidates using CRISPR/Cas9 gene targeting in zebrafish embryos. (a) Sanger sequence traces for genomic DNA PCR analysis of CRISPR/Cas9 targeted embryos show successful genetic disruption at the sgRNA site loci for eph3ba, eph3bb, and emx2. PAM sites are indicated. (b) Brightfield microscopy analysis of targeted fish at 3 dpf and 5 dpf compared to uninjected controls and Cas9-only controls. (c) Eye diameter metrics for fish injected with CRISPR/Cas9 at 3 dpf and 5 dpf (**** = p < 0.0001). (d) Confocal analysis of laminin-stained cryosections from gene-edited and control eyes showing persistence of the basement membrane in the optic fissure margin (arrowheads). Colobomas were observed in 27% and 47% of gene-edited embryos analysed for emx2 and ephb3a/b, respectively (n = 15 per target gene).

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Acknowledgments

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