Fig. 1: Zebrafish models with setd2 loss-of-function mutations are viable. | Cell Discovery

Fig. 1: Zebrafish models with setd2 loss-of-function mutations are viable.

From: setd2 knockout zebrafish is viable and fertile: differential and developmental stress-related requirements for Setd2 and histone H3K36 trimethylation in different vertebrate animals

Fig. 1

a Schematic diagram showing ___domain architecture of zebrafish Setd2 protein and mutated sites of the three mutant lines generated by CRISPR/Cas9 genome editing. The wild-type (WT) setd2 mRNA is referred to the NCBI Reference Sequence XM_009292184.3. Two mutants contain frame-shift mutations in exon 3 and are therefore named E3fs1 (c.215G>A216_217del; p.R72Qfs*9) and E3fs2 (c.217_244del; p.Q73fs*133), both of which disrupt the majority of the protein. The third mutant contains a frame-shift in exon 19, named E19fs1 (c.7910_7913GTGA>TTTC7914del; p.S2637Ifs*34), which disrupts the C-terminal SRI ___domain. Genomic sequences and deduced amino acids around targeting sequences in setd2 are shown. Red letters denote sgRNA-targeting sequences, while blue letters and dotted lines represent mutant sequences. b–d Genotype ratios of the WT, heterozygous (HE) and homozygous (HO) progenies, which are produced by self-cross of the three mutants, and their persistence through growth into adult. Note that they all fit the Mendelian Ratio. e Immunoblot analysis of the different H3K36 methylation states in the WT, HE and HO embryos of the E3fs1 and E19fs1 mutants. Total H3 was used as a loading control. Embryos at 72 hpf were analyzed. Each sample was loaded in 2-fold serial dilution to facilitate quantification. The immunoblot of the E3fs1 samples was actually analyzed in the same gels/explorations, but the images were rearranged just to fit the loading order in the lower panel. f Quantification of H3K36me3 levels in HE and HO relative to WT embryos of the E3fs1 and E19fs1 mutants.

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