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Figure 5
(A) Unmodified, TEG, and 2?-OMe-RNA::TEG-modified double-stranded DNA (dsDNA) donors were injected into zebrafish zygotes. dsDNA donor design to knock-in Avi-tag is shown on the top and the fraction of Illumina reads containing precise knock-in are plotted as percentages. Mean ± s.d. for at least three independent replicates (two for unmodified donors) are plotted (B). Design of the dsDNA donors injected into mouse zygotes to precisely convert the coat color of albino mice (TyrC) to pigmented (TyrC-Cor) by editing C to G (underscored) along with six silent mutations (in red) is shown. Percentages of F0 founder mice with black coat are shown. (C) Percentages of animals among homology-directed repair (HDR)-positive F0s that have uniform dark coat or mosaic coat color are plotted for unmodified and 5?-modified donors. (D) Representative pictures of 10-day- old F0 mice with pigmented (HDR) or white (wild-type [WT] or indel) coat color are shown. One mosaic mouse (third from left) can be seen among the pups obtained with end-modified donor. (E) Donor design to knock-in V5 tag at the C-terminus of Sox2 is shown on the top. Percentage of founder animals containing perfect V5 insertion at Sox2 locus are shown for each donor type. HA, homology arms. p-Values were calculated using one-way ANOVA (Tukey?s multiple comparisons test; ****p < 0.0001; ***p < 0.001; **p < 0.01; *p < 0.05; ns, not significant).
2?-O-methyl (2?-OMe)-RNA-triethylene glycol (TEG) donors promote precise editing in vertebrate zygotes.