FIGURE

Fig. 1

ID

ZDB-FIG-220609-1

Publication

Zhang et al., 2021 - Rapid generation of maternal mutants via oocyte transgenic expression of CRISPR-Cas9 and sgRNAs in zebrafish

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Fig. 1

(A) Construction of pGGDestISceIEG-1sgRNA plasmid that contains a single sgRNA expression cassette and a GFP expression module driven by the ef1α promoter. I-SceI restriction sites and TIR are designed to flank sgRNA and GFP sequences for transgenesis. EGFP, enhanced GFP. (B) Construction of the transgenic pGGDestISceIEG-3sgRNA vector expressing three tandem sgRNAs via Golden Gate ligation. (C) Pipeline to generate maternal mutants in F1 embryos. The sgRNA expression cassette is introduced into Tg(zpc:zcas9) embryos by I-SceI–mediated transgenesis, and the phenotype of the resulting GFP-positive F1 embryos is examined to identify developmental defects. CmR represents the chloramphenicol resistant gene, while EG highlighted in green designates the GFP expression cassette driven by the ef1α promoter. WT, wild type.

Expression Data

Expression Detail

Antibody Labeling

Phenotype Data

Phenotype Detail

Acknowledgments

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