FIGURE Generation of fbn1 ^+/? zebrafish. The conserved domains of FBN1 protein in zebrafish are shown. The red box and the red arrow indicate the target of CRISPR/Cas9 gene editing. The CRISPR/Cas9 induced a frameshift mutation containing 3?base deletion and 16?base insertion in fbn1. The inserted nucleotides are marked in red. The red dashed line indicates the 3?base deletion. WT: wildtype; TB: transforming growth factor?beta?binding protein?like

FIGURE Identification of F2 fbn1 ^+/? zebrafish. (a) PCR amplification of the genomic DNA from TU wild?type and F1 fbn1 ^+/? zebrafish. M, DNA marker. (b?d) PCR amplification of the genomic DNA of F2 fbn1 ^+/? adults. Lanes 9 and 26, DNA Marker; lanes 3, 4, 6, 8, 12?14, 17, 18?20, 22, 25, 27, 29?32, and 34 in (b), heterozygous for alleles A and B mutants; lanes 1, 2, 5, 7, 9?11, 15, 16, 21, 23, 24, 26, 28, and 33 in (b), wild?type; lanes 1?3, 10?12, 14?16 in (c), heterozygous mutants of alleles A and B; lanes 4?9, 13, and 17 in (c), wild?type; lanes 1, 3, 4, and 7 in (D), heterozygous mutants of alleles A and B; lanes 2, 5, 6, 8, and 9 in (d), wild?type. (e) DNA sequencing results of F2 fbn1^+/? fish. Red arrow indicates the position of the mutation

(a) Represents the ?family portrait? of the F2 fbn1 ^+/? zebrafish (experimental group) generated by the hybridization of F1 fbn1 ^+/? heterozygous mutants and Tg(cmlc2:eGFP) transgenic fluorescent zebrafish. The picture on the left is the morphology of F2 fbn1 ^+/? zebrafish at 72 hr post?fertilization, and the picture on the right is a 120?hr ?family portrait? of F2 fbn1 ^+/? zebrafish. In (b) group a is the fluorescence micrograph of TU wild?type embryos, and group b is the fluorescence micrograph of F2 embryos (experimental group), where a1 and b1 are 72 hr fluorescence images, a2 and b2 are 120 hr fluorescence images. The results showed that no significant difference was found in the early developmental observation between the TU wild?type and F2 generation experimental groups. The legend has been marked with the meaning of "a1, a2, b1, b2." Scale bars have been marked in Figure ?Figure3.3. Magnification eyepiece ×0.6 (A) and eyepiece ×2.0 (B). hpf: hours post?fertilization

Comparison of the morphology, head, and heart between F2 fbn1 ^+/? and TU wild?type zebrafish larvae. Representative images of the morphology of F1 and F2 fbn1 ^+/? zebrafish at 12, 19, 26, 33, and 40 days post?fertilization are shown. F2 fbn1 ^+/?zebrafish exhibited more noticeably decreased melanin, the red arrow in the figure refers to the typical representatives of F2 embryos showing the characteristics of markedly reduced melanin at the 12 days post?fertilization. Scale bars have been marked in Figure ?Figure4.4. Magnification eyepiece ×0.6 (left), ×1.0?2.0 (middle), and ×3.5 (right). dpf: days post?fertilization