Fig. 4

Fig. 4

Injection of the predicted MMEJ-inducing dnajb6b-sgRNA into bag3^e2/e2 F0 fish manifested severe cardiac dysfunction. (A) Design of the predicted MMEJ-based sgRNA sequence targeting the exon 6 of the dnajb6b gene (dnajb6b^e6-MJ). Injection of dnajb6b^e6-MJ into bag3^e2/e2 fish (bag3^e2/e2;dnajb6b^e6−MJ) generated frame-shift mutant alleles, with 37.5% predicted to result from microhomology recombination (7-nucleotide deletion). (B) Plotted is the dnajb6b gene knockout (KO) score (in %) of bag3^e2/e2;dnajb6b^e6−MJ embryos at 3 dpf and fish at 3 months. (C) Transcript levels of dnajb6b in hearts of bag3^e2/e2;dnajb6b^e6−MJ fish compared to those of non-injected controls at 3 months; n=10. Unpaired Student’s t-test. (D,E) Fraction shortening (FS) and ejection fraction (EF) in bag3^e2/e2;dnajb6b^e6−MJ fish compared to that in bag3^e2/e2 mutant, dnajb6b^e6-MJ or non-injected WT control fish at 6 dpf (embryonic) and 6 months (adult). n=5-8, one-way ANOVA.