ZFIN Image: Diofano et al., 2020, Fig 1

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Figure Caption

Fig 1 Generation of zebrafish <italic>bag3</italic> knockout by CRISPR/Cas9 gene editing.

(A) Structure of the zebrafish bag3 gene and protein. Exon 2 is the target for the CRISPR/Cas9 gene editing in zebrafish bag3. The CRISPR/Cas9-induced mutation (19 bp deletion) in bag3 is shown in bag3 mutant DNA sequencing chromatogram. The 19 nucleotides deletion in bag3^-/- leads to a frame shift, the introduction of a premature stop codon and thereby the premature termination of Bag3 translation, as demonstrated by the alignment of the Bag3^-/- and Bag3^+/+ aminoacid sequences (only partial aminoacid sequence shown) (B-C) Immunoblot analysis of 72 hpf bag3^+/+ embryo protein lysates compared to lysates obtained from bag3^-/- clutchmates with antibody against zebrafish Bag3. The figure shows one representative immunoblot from three independent experiments (N = 3, mean ± SD, P<0.0001 determined using two-tailed t-test). (D) Quantitative real-time PCR of bag3^+/+ and bag3^-/- embryos at 72 hpf shows significant downregulation of bag3 mRNA levels in bag3^-/- embryos (N = 3, mean ± SD, P = 0.0004 determined using two-tailed t-test).

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Acknowledgments

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