Fig. 5

(A) Representative ventral images of −1.4col1a1:egfp zebrafish larvae at 3 days post-fertilization (dpf). Orientation arrows indicate anterior (A), posterior (P), left (L), and right (R). Scale bar, 300 μm.

(B) Quantitative assessment of the CHA of larvae injected with single human mRNAs for each of the 30 genes located in the 16p11.2 BP4-BP5 region. Images were measured as shown in (A) (angle between dashed lines). Seven transcripts induced a significant reduction in CHA after Tukey’s p value adjustment (adjusted p < 0.01). Dosage is 12.5 pg for KIF22 and PPP4C and 50 pg for all other genes.

(C) Quantitative assessment of the CHA of F0 mutant batches injected with single combinations of each of sez6l2, taok2a, and taok2b gRNAs with or without Cas9. Dosage is 50 pg gRNA and 200 pg Cas9 protein.

(D) Quantitative assessment of the CHA of larvae injected with single or equimolar combinations of human KCTD13, MAPK3, and MVP mRNAs. Dosage is 50 pg.

(E) Quantitative assessment of the CHA of F0 mutant batches injected with single or equimolar combinations of kctd13, mapk3, and mvp gRNAs with or without Cas9. Dosage is 50 pg gRNA and 200 pg Cas9 protein. The number of larvae measured for each condition is indicated at the base of each bar in the graphs. The data are represented as the mean ± SEM; ns, not significant; **p < 0.01, ***p < 0.001, and ****p < 0.0001 versus uninjected controls. Tukey’s multiple comparison tests were applied following a significant one-way ANOVA.