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. 2017 Nov 29;208(2):673–686. doi: 10.1534/genetics.117.300468

Figure 2.

Figure 2

slc45a2 knockout results in loss of eye and body pigmentation and can efficiently be targeted to the eye. (A) Phenotype severity scoring system based on pigment loss in the eye. Mild phenotype: >50% of eye pigment present; moderate phenotype: <50% of eye pigment present; severe phenotype: almost no eye pigment present. Embryos injected with 500 pg slc45a2 sgRNA, 1 ng Cas9 protein, and memRFP RNA tracer in one cell of a 2-cell embryo. (B) Survival and phenotype severity of embryos injected with varying amounts of slc45a2 sgRNA and Cas9 protein. Exogastrulation: embryo death prior to gastrulation; death: embryo death after gastrulation and before stage 39–41; none: no eye pigment loss. Surviving stage 39–41 embryos were scored based on phenotype severity. (C) Phenotype severity eye-targeted CRISPR gene editing. Embryos injected with 500 pg slc45a2 sgRNA, 1 ng Cas9 protein, and Alexa 488 dextran tracer at 1-, 2-, and 8-cell stages (blastomere D1) show slc45a2 editing in the targeted eye tissue. Embryos shown at neurula (NF stage 19–20) and tadpole (NF stage 39–41) stages. Mistargeted: tracer not present in the eye. (A and C) White bar, 500 µm, black bar, 100 µm. * Injected side of embryo.