Generation of Albino Cynops pyrrhogaster by Genomic Editing of the tyrosinase Gene.
Albino animals are useful for in situ hybridization experiments that demonstrate gene expression in embryos and organs, for the immunological rejection of skin grafts transplanted to host animals, and to identify tissues with regenerative ability during limbs and retina regeneration processes. Cynops pyrrhogaster has extensive regenerating capacities. To facilitate regenerative research, in the present study, we produced albino C. pyrrhogaster using genomic editing. The DNA fragment containing part of the tyrosinase gene from C. pyrrhogaster was amplified using degenerate primers corresponding to evolutionarily conserved nucleotide sequences among several species, and the nucleotide sequence was determined. We designed a transcription activator-like effector nuclease (TALEN) that targets a candidate of the C. pyrrhogaster tyrosinase gene. Fertilized eggs were injected with TALEN mRNA, and albinos of C. pyrrhogaster were obtained. The results of the present study demonstrated that TALEN can be used effectively for genomic editing in C. pyrrhogaster and that the candidates of the tyrosinase gene that were cloned by us are essential for melanin synthesis. The albino newts created in the present study can be used as versatile experimental material.
Keisuke
K
Nakajima
Division of Embryology and Genetics, Institute for Amphibian Biology, Graduate School of Science, Hiroshima University, 1-3-1, Kagamiyama, Higashihiroshima, Hiroshima 739-8526, Japan.
Taeko
T
Nakajima
Division of Embryology and Genetics, Institute for Amphibian Biology, Graduate School of Science, Hiroshima University, 1-3-1, Kagamiyama, Higashihiroshima, Hiroshima 739-8526, Japan.
Yoshio
Y
Yaoita
D000818:Animals; D001483:Base Sequence; D004252:DNA Mutational Analysis; D000072669:Gene Editing; D005786:Gene Expression Regulation / Q000235:genetics*; D008543:Melanins / Q000235:genetics*; D014442:Monophenol Monooxygenase / Q000737:chemistry / Q000235:genetics*; D012455:Salamandridae / Q000235:genetics*; D016415:Sequence Alignment; D000069896:Transcription Activator-Like Effector Nucleases / Q000378:metabolism; D015053:Zygote / Q000502:physiology
D008543:Melanins; D014442:Monophenol Monooxygenase; D000069896:Transcription Activator-Like Effector Nucleases
Cynops pyrrhogaster; TALENs; albino phenotype; genomic editing; targeted gene knockout; tyrosinase