Figure 1

Impact of targeting the setb gene on Hedgehog signaling in zebrafish embryos. (A) Zebrafish setb crispants exhibit cyclopia. One- to two-cell stage zebrafish embryos were injected with 4.6 nl of 150 ng/μl of Cas9 mRNA or with 50 ng/μl of setb gRNA and 150 ng/μl of Cas9 mRNA and were examined morphologically at 48 hpf. (Right graph). Phenotypic scoring of the injected embryos (n = 262, n denotes the number of injected embryos). Numbers in X-axis refer to percentage of embryos in each phenotypic group and numbers on the bars refer to the percentage of cyclopic embryos in each phenotypic group. (B) Hh signaling is reduced in Tg12x_Gli transgenic zebrafish embryos injected with setb gRNA/Cas9. One- to two-cell stage Tg12x_Gli transgenic zebrafish embryos were injected with setb gRNA/Cas9 and examined under a confocal fluorescence microscope at 24 hpf. The phenotypic scoring of the injected embryos is shown in the graph (n = 96). The squares show enlargement of the trunk areas that display decreased mCherry fluorescence intensity. Note the disrupted morphology of the somites in the marked regions.