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Fig. S1 J
Fabp11a gene knockout using CRISPR/Cas9 system caused eye developmental defects in zebrafish. (A) Schematic diagram showing Guide RNA targeting sites on the exons of fabp11a gene. Guide DNA targeting sites are indicated by arrowheads. The targeting sequences are highlighted in orange and purple. (B) Graphical view of Fabp11a protein domain. (C) Sanger sequencing analysis of PCR fragments amplified from gRNA1 and gRNA2 target regions in gRNA1/gRNA2 and cas9 mRNA coinjected embryos. (D) Mutation pattern of gRNA1/gRNA2 and cas9 mRNA coinjected embryos. Numbers in the brackets show the number of nucleotides were deleted (-) or inserted (+). Inserted nucleotide is in red. WT, wild type. (E) Microscopy analysis of eyes in control and gRNA1-cas9 mRNA coinjected embryos, dorsal view. Red arrowheads indicate normal paired eyes. Blue arrowhead indicates no eye in gRNA1-cas9 mRNA coinjected embryo. Green arrowhead indicates single fused big eyes, dorsal view. Purple arrowhead indicates single eye, dorsal view. (F) Statistical analysis of phenotype efficiency in control and gRNA1-cas9 mRNA coinjected embryos. χ² test; ****, P<0.0001. (G) Microscopy analysis of eyes in control and fabp11a-/- embryos. Arrowheads indicate eyes.