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Optic cup architecture and size quantification in candidate gene crispant embryos.a–o Representative optical sections through the optic cup of crispant 24 hpf embryos. Phalloidin/DAPI staining reveals tissue organization of control embryos injected only with Cas9 (a), with sgRNAs directed against rx3 (b), or with sgRNAs for the different candidate genes (indicated in (c)–(o)). Note that, in contrast to rx3 crispant embryos, the optic cup is formed in all cases. However, quantitative analysis of eye area (p) reveals a significant reduction of optic cup size for all candidates tested with the exception of nr2f1. Individual values (n = 10) are plotted in front of standard box-and-whiskers (Two-tailed T-test); ***=p < 0.001, ****=p < 0.0001. p values: cas9 = 0.161; rx3 = 1.15e−6; crh1a = 8.67e−5; dsp = 1.82e−4; mcm2 = 3.09e−5; mif = 1.05e-9; neurod4 = 9.00e−8; nop58 = 8.09e−6; nr2f1 = 0.053; smad6b = 2.25e−4; tcf12 = 2.71e−9; tead1 = 1.62e−6; tead1 + 3 = 1.15e−6; vgll2 = 9.15e−7; wdr12 = 5.00e−4. Abnormal morphology of the RPE cells (m,n) is indicated (red arrowheads). ov optic vesicle, lv lens vesicle. Bar = 50 µm. Source data are provided as a Source Data file. See also Supplementary Figs. 9 and 10.
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