Fig. 2

Fig. 2

Generation of creb3l3a and creb3l3b zebrafish mutants. A: Representative images of whole-mount in situ hybridization (ISH) with sense and antisense probes targeting zebrafish creb3l3a (ENSDARG00000056226.7) and creb3l3b (ENSDARG00000090903.3) at 6 days post fertilization (dpf). Both creb3l3a and creb3l3b mRNA is expressed in the liver and the intestine at 6 dpf (white outlines). No signal is observed with sense probes. Images are representative of 3 independent clutches: 6–12 animals per clutch. B: Overview of gene loci and strategy used for CRISPR/Cas9 targeting of creb3l3a and creb3l3b. The fifth exon was targeted by CRISPR guide RNA in creb3l3a, and the sixth exon was targeted in creb3l3b. C: Sanger sequencing confirms 2-bp deletions in both creb3l3a and creb3l3b (nucleotides depicted in gray show the affected codon causing a frameshift; dotted box around sequence trace confirms codon change by loss of 2 base pairs). Creb3l3, cAMP-responsive element-binding protein 3–like 3.