Figure 2

Figure 2

Generation of cfdp1 zebrafish mutant line. (A) Schematic representation of the zebrafish cfdp1 gene. For generation of the CRISPR/Cas9-mediated mutant line, a target site in exon 3 was selected. (B) Schematic representation of the strategy for CRISPR/Cas9-mediated zebrafish line. The injection mix of gRNA/Cas9 was injected at the one-cell-stage embryos. The crispants (F0-injected) grow until adulthood and are crossed with wild-type adults. The F1 generation is genotyped to identify possible founders of the line. After the identification, the corresponding F1 heterozygous generation is kept for further analysis. (C) Upper: nucleotide alignment between cfdp1 mutant and cfdp1 wild-type sequence. A 5 bp deletion in cfdp1 mutant is detected. Lower: chromatogram of Sanger sequencing of cfdp1 mutant and cfdp1 wild-type sequence and the corresponding aa they encode. In the cfdp1 mutant, at the point of DNA break, seven novel amino acids (aa) are inserted before a premature stop codon.