Figure 1.

Schematic illustration of TALEN-mediated knockout of the zebrafish Prkdc gene. (A) Generation of TALEN constructs. Exon 3 of the Prkdc gene was targeted with the 12-bp spacer flanked by left and right TALEN arms. (B) Mutagenesis by microinjection of capped mRNA and strategy for genotyping of the F1 progenies. (C) Sequence confirmation of the mutations with two-nucleotide deletion and six-nucleotide deletion followed by five-nucleotide insertion (mutation no. 3 and 6, respectively). Both mutations cause a shift of the translational frame. (D) Western blot analysis for Prkdc. Kidney and spleen were dissected from several zebrafish, and the pooled samples were processed for the Western blot experiment. C, Prkdc^+/−; P, Prkdc^−/−. (E) IHC experiment for Prkdc done with the kidney dissections.