Fig. 1

A Site-specific targeting for CRISPR/Cas9 cleavage within exon 12 of the zebrafish asxl1 gene. Alignment of nucleotide sequences from wild-type and mutant asxl1 alleles in asxl1^{e12 (−7)} and asxl1^{e12 (−22)} zebrafish lines. Dashes in DNA sequences are the nucleotides deleted during repair of CRISPR/Cas9-induced double-strand breaks. PAM protospacer adjacent motif. CRISPR/cas9-induced asxl1 frameshift mutations predicted to lead to C-terminally truncated proteins. Truncated proteins predicted from mutant alleles asxl1^{e12 (−7)} and asxl1^{e12 (−22)} lack the last two domains (ASXM2 and PHD). B qRT-PCR comparing expression of asxl1 in asxl1^+/+ and mutant asxl1^−/− in 3 days postfertilization (dpf) larvae and adult kidney marrow (3 dpf, 3 dpf larvae tails, n ≥ 10 per group, performed with four replicates; 6 m, 6-month kidney marrow, n = 4 per genotype; two-tailed Student’s t test, *p < 0.05, **p < 0.01, ***p < 0.001; error bars, mean ± standard deviation (SD)). C Gross appearance of asxl1^−/− zebrafish compared with asxl1^+/+ littermates (at 1.5 years). Body weights of asxl1^+/+ and asxl1^−/− zebrafish (scale bar, 1 cm; asxl1^+/+, n = 33; asxl1^−/−, n = 24; two-tailed Student’s t test, *p < 0.05, **p < 0.01, ***p < 0.001, error bars, mean ± SD).