Fig. 1
- ID
- ZDB-FIG-220617-38
- Publication
- Rosello et al., 2022 - Disease modeling by efficient genome editing using a near PAM-less base editor in vivo
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Efficient base conversion generates albino F0 embryos with CBE4max-SpRY and a NAN PAM.
a Targeted genomic sequence of the tyrosinase gene and the 3 different sgRNAs used to introduce the W273* mutation. b Targeted genomic sequence of the slc45a2 gene and the 2 different sgRNAs used to introduce the W121* or R123* mutations. a, b. For each sgRNA, the targeted Cs are in red and the PAM sequence is in green. c Lateral view of representative 2 days post-fertilization (dpf) embryos showing different severity of pigmentation defects (wild-type like, mildly depigmented, severely depigmented, and albino embryos). Scale bar = 100 µm. d Proportion of the 4 groups based on the pigmentation defects described in Fig. 1c for each injection: the AncBE4max mRNA and the tyr(W273*)NGG sgRNA (column 2, 19 embryos in total), the CBE4max-SpRY mRNA and the tyr(W273*)NGG sgRNA (column 3, 74 embryos in total), the CBE4max-SpRY mRNA and the tyr(W273*)NAN sgRNA (column 4, 28 embryos in total), the CBE4max-SpRY mRNA and the tyr(W273*)NGN sgRNA (column 5, 10 embryos in total). e Proportion of the 4 groups based on the pigmentation defects described in c for each injection: the CBE4max-SpRY mRNA with the slc45a2(R123*) sgRNA (column 2, 100 embryos in total) or with the slc45a2(R121*) sgRNA (column 3, 126 embryos in total). f C-to-T conversion efficiency for each targeted Cs and each pool of embryos showing pigmentation defects presented in d, e. The efficiencies have been calculated using EditR tool and chromatograms from Sanger sequencing made on entire PCR products. |