Fig. 4

Comparing mRNA rescue levels of OGDHL and OGDH in Dhtkd1 and Ogdh paralog-deficient zebrafish. A Representative image of uninjected, Cas9-injected, single knockout (dhtkd1, ogdha, and ogdhb F₀), double knockout (ogdha;ogdhb F₀), and triple knockout (ogdha;ogdhb;ogdhl F₀) embryos at 3 dpf. B, C Size measurements for head and eye with or without human OGDHL co-injection. The number of embryos for uninjected = 66, Cas9-injected = 60, dhtkd1 F₀ = 36, dhtkd1 F₀ + OGDHL = 36, ogdha F₀ = 66, ogdha F₀ + OGDHL = 36, ogdhb F₀ = 66, ogdhb F₀ + OGDHL = 36, ogdha;ogdhb F₀ = 36, ogdha;ogdhb F₀ + OGDHL = 36, ogdha;ogdhb;ogdhl F₀ = 36 and ogdha;ogdhb;ogdhl F₀ + OGDHL = 36. D, E The size measurements for head and eye with or without human OGDH co-injection. The number of embryos for uninjected = 60, dhtkd1 F₀ = 36, dhtkd1 F₀ + OGDH = 36, ogdha F₀ = 36, ogdha F₀ + OGDH = 36, ogdhb F₀ = 36, ogdhb F₀ + OGDH = 36, ogdhl F₀ = 36, and ogdhl F₀ + OGDH = 36. All values were calculated as a percentage difference compared to the mean value of the uninjected embryos. Each dot represents one embryo, and the mean value of each group is indicated at the bottom of the respective bar in the figure. Error bars = mean ± SD. Statistical significance was calculated by Brown–Forsthye and Welch’s ANOVA with Dunnett’s T3 multiple comparisons test: not significant (ns) p ≥ 0.05, *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001