XB-IMG-152306
Xenbase Image ID: 152306
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Figure 1. Targeted gene disruption of tyrosinase (tyr), solute carrier family 45 member 2 (slc45a2) and leukocyte tyrosine kinase (ltk) using Cas9 protein or Cas9 mRNA with sgRNA in Xenopus tropicalis founders. sgRNA target protospacer sequences are shown by pink letters, and protospacer-adjacent motif (PAM) sequences are shown in red boxes. (A) Disruption of tyr in founders. (Left) Representative phenotypes of tyr-disrupted embryos. Phenotypes were classified into four groups: severe, near complete loss of pigmentation in retinal pigmented epithelium (RPE); moderate, more than half of pigmentation lost; weak, less than half of pigmentation lost; and normal, no alteration in pigmentation. (Right) Frequencies of tyr-disrupted phenotypes. Injections of Cas9 protein with tyr sgRNA in buffer A or B were independently conducted three and two times, respectively. Injection of Cas9 mRNA with tyr sgRNA was independently carried out three times. (B) Disruption of slc45a2 in founders. (Left) Representative phenotypes of slc45a2-disrupted embryos using Cas9 mRNA and sgRNA. Phenotypes were classified into four groups as described for tyr phenotypes. Severe phenotype was near complete pigmentation loss. (Right) Frequencies of slc45a2-disrupted phenotypes from three independent experiments. (C) Disruption of ltk in founders. (Left) Representative phenotypes of ltk-disrupted tadpoles using Cas9 mRNA and sgRNA. Phenotypes were classified into three groups: full, the loss of iridescence in both eyes; half, the loss of iridescence in either eye; and normal, no alteration in iridescence in both eyes. (Right) Frequencies of ltk-disrupted phenotypes from two independent experiments. Total numbers of individuals are shown at the top of each graph. In vitro fertilization (IVF) indicates control embryos without injection Image published in: Shigeta M et al. (2016) Copyright © 2016. Image reproduced with permission of the Publisher, John Wiley & Sons.
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