|
Figure 1. pdip expression is undetectable in X. tropicalis ptf1a/p48-null mutant tadpoles. Whole-mount in situ hybridization analysis of pdip expression in F2 X. tropicalis ptf1a/p48 mutant tadpoles (stage 43). Ventral-lateral view, head to the left. (A, C) About 22% homozygous tadpoles showed complete absence of pdip expression. (B, D) The remainder uniformly showed strong pdip expression, suggesting normal pancreas development in ptf1a/p48 heterozygous tadpoles.
|
|
Figure 2. CRISPR/Cas9 is effective in targeting X. laevis ptf1a/p48. DNA sequencing data reveal the indel-inducing efficiencies of ptf1a/p48-T1 and ptf1a/p48-T2 in X. laevis embryos. For all panels, the wild-type sequence is shown at the top with the target site highlighted in yellow and the PAM sequence in blue. Red dashes indicate deletions (−) and lowercase letters in red indicate insertions (+).
|
|
Figure 3.
ptf1a/p48-targeted X. laevis G0 tadpoles showed reduction of pdip expression. Whole-mount in situ hybridization analyses of pdip expression (stage 42). Ventral-lateral view, head to the left. (A, B) Wild type X. laevis tadpoles. (C, E) Severe reduction. (D, F) Mild reduction. (G, I) Severe reduction. (H, J) Mild reduction. (K, N) Almost complete absence. (L, O) Severe reduction. (M, P) Mild reduction.
|
|
Figure 4. CRISPR/Cas9 is effective in targeting X. laevis tyrosinase. DNA sequencing data reveal the indel-inducing efficiencies of tyra-T and tyrb-T in X. laevis embryos. For all panels, the wild-type sequence is shown at the top with the target site highlighted in yellow and the PAM sequence in blue. Red dashes indicate deletions (−) and lowercase letters in red indicate insertions (+). The left two panels show data from tyra-T and tyrb-T co-injection and right two panels indicate individual injection results.
|
|
Figure 5. Simultaneous disruption of two X. laevis tyrosinase homeologs leads to albinism in G0 tadpoles (stage 42). Lateral view, head to the left. (A, D) Almost full albinism. (B, E) Severe inhibition of pigmentation. (C, F) Almost normal. (G, J) Wild type. (H, I, K, L) Almost normal.
|
|
pdia2 (protein disulfide isomerase family A member 2 ) gene expression in X. laevis embryos, assayed via in situ hybridization, NF stage 43, lateral view, anterior left, dorsal up.
|