Fig. 3.

Generation of large deletion of TYR in rabbits by dual sgRNAs. a Phenotype of TYR mutant rabbit; #301 is the TYR biallelic mutant (homozygote), which exhibited the typical albinism phenotype; #204 and #106 are the TYR monoalleic mutation (heterozygote) and the wild-type rabbit. The box shows the light pink-tinted iris and pupil of the #301 rabbit compared with the dark iris of wild-type rabbits. WT wild-type control. b The mutation determination of 105 kb deletions of TYR gene in founder rabbits by PCR. Results showed that three (#106, #204 and #301) of the 17 founders were large deletion of TYR rabbit. Primers used were TYR-F1 and R1 (Table S1). M DNA marker. c T-cloning sequences of mutant alleles in the large deletion of TYR rabbit (#106, #204 and #301). The PAM sites are underlined and highlighted in red; the target sequences are green; deletions (−) and insertions (+) are shown. WT wild-type control. d Chimera analysis of different tissues from #204 by PCR-sequencing. All detected tissues showed the same PCR band. Primers used were F1 and R1 (Table S1). M DNA marker, WT wild-type control. e T-cloning sequences of mutant alleles in different tissues from #204. PAM sites are underlined and highlighted in red; target sequences are green; deletions (−) and insertions (+) are shown. WT wild-type control