Double knock-out of pax6 and tyr in Xenopus F0 embryos.

(A) Double knock-out of pax6 and tyr was achieved by TALEN mRNA injection to GV oocytes. A double knock-out tadpole is shown. (B) Detection of pax6 and tyr mutants by RFLP analysis. Wild type embryos are cut by restriction enzymes (Ui: uninjected embryo), while mutant embryos are not digested. Digested PCR products appear at lower bands (marked by open arrowheads) and the undigested is marked by closed arrowheads. M represents 100 bp ladder marker. (C) Sequencing of three different embryos (#3, #6, #9; the numbers correspond to those in Fig 3B) revealed the 100% mutation rate in pax6 and tyr double knock-out embryos.