FIGURE SUMMARY
Title

Evaluation and application of modularly assembled zinc-finger nucleases in zebrafish

Authors
Zhu, C., Smith, T., McNulty, J., Rayla, A.L., Lakshmanan, A., Siekmann, A.F., Buffardi, M., Meng, X., Shin, J., Padmanabhan, A., Cifuentes, D., Giraldez, A.J., Look, A.T., Epstein, J.A., Lawson, N.D., and Wolfe, S.A.
Source
Full text @ Development

The ZFN-induced um27 lesion is a truncation allele of gata2a. (A) Location of frameshift caused by the um27 deletion in the zebrafish gata2a gene. ZF, zinc-finger domain. (B) gata2a coding and amino acid sequence in the region of the um27 deletion. ZFN recognition sequences are boxed. (C) Genotype of embryos derived from an incross of gata2aum27 heterozygous carriers. (D) Genotype of embryos with normal circulation derived from a gata2aum27 incross injected with 200 pg gata2a mRNA. Asterisks denote mutant embryos with normal trunk circulation. (E) Percentage of gata2aum27 mutant embryos displaying the indicated circulation phenotypes.

PHENOTYPE:
Fish:
Observed In:
Stage: Long-pec

gata2aum27 mutant embryos display defects in aorta morphogenesis. Confocal microangiography using Quantum Dots in Tg(kdrl:egfp)la116 transgenic zebrafish embryos. Endothelial cells are green and vessel perfusion is red. (A,B) Cranial blood vessels in gata2aum27 mutant (A) and wild-type (B) embryos. Lateral views, anterior to the left, dorsal is up. (C,D) Lateral dorsal aortae (arrows) in gata2aum27 mutant (C) and wild-type (D) embryos. Dorsal views, anterior is up. (E,F) Trunk blood vessels in gata2aum27 mutant (E) and wild-type (F) embryos. Lateral views, anterior to the left, dorsal is up. Segmental vessels are indicated by arrowheads and the dorsal aorta by an arrow; the asterisk indicates a region of aorta that failed to form.

gata2aum27 mutant embryos show reduced kdrl expression but normal artery and blood differentiation. (A-F) Whole-mount in situ hybridization at 24 hpf using antisense riboprobes against hey2 (A,B), tal1 (C,D) and kdrl (E,F) in genotypically wild-type zebrafish embryos (A,C,E) and those homozygous for the um27 deletion (B,D,F). Lateral views, anterior to the left, dorsal is up.

Morphology of wild-type and gata2aθm27 mutant embryos. (A-F) Wild-type sibling (A-C) and gata2aum27 mutant (D-F) zebrafish embryos at 48 hpf (A,D) and 24 hpf (B,C,E,F). (A,D) Images captured on a standard dissection microscope. (B,C,E,F) Differential interference contrast images. nc, notochord; nt, neural tube; DA, dorsal aorta; PCV, posterior cardinal vein. Arrows indicate somite boundaries and arrowheads denote horizontal myoseptum. All images are lateral views, anterior is to the left, dorsal is up.

PHENOTYPE:
Fish:
Observed In:
Stage Range: Prim-5 to Long-pec

Unillustrated author statements

Acknowledgments
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