title: Generation of myostatin B knockout yellow catfish (Tachysurus fulvidraco) using transcription activator-like effector nucleases.
abstract: Myostatin (Mstn), a member of the transforming growth factor β superfamily, plays an inhibiting role in mammalian muscle growth. Mammals like human, cattle, mouse, sheep, and dog carrying null alleles of Mstn display a double-muscle phenotype. Mstn is conserved in fish; however, little is known whether the fish with mutated mstn display a similar phenotype to mammals because of the lack of mutant fish with mstn null alleles. Previously, we knocked out one of the duplicated copies of myostatin gene (mstna) in yellow catfish using zinc-finger nucleases. In this study, we report the identification of the second myostatin gene (mstnb) and knockout of mstnb in yellow catfish. The gene comprises three exons. It is predicted to encode 373 amino acid residues. The predicted protein exhibits 59.3% identity with yellow catfish Mstna and 57.3% identity with human MSTN. Employing TALEN (transcription activator-like effector nucleases) technology, we obtained two founders (from four randomly selected founders) of yellow catfish carrying the mutated mstnb gene in their germ cells. Totally, six mutated alleles of mstnb were obtained from the founders. Among the six alleles, four are nonframeshift and two are frameshift mutation. The frameshift mutated alleles include mstnb(nju22), an 8 bp deletion, and mstnb(nju24), a complex type of mutation comprising a 7 bp deletion and a 12 bp insertion. They are predicted to encode function null Mstnb. Our results will help to understand the roles of mstn genes in fish growth.
authors.affiliation: 1 MOE Key Laboratory of Model Animal for Disease Study, Model Animal Research Center, Nanjing University , Nanjing, China .
authors.forename: Zhangji
authors.initials: Z
authors.lastname: Dong
authors.forename: Jiachun
authors.initials: J
authors.lastname: Ge
authors.forename: Zhiqiang
authors.initials: Z
authors.lastname: Xu
authors.forename: Xiaohua
authors.initials: X
authors.lastname: Dong
authors.forename: Shasha
authors.initials: S
authors.lastname: Cao
authors.forename: Jianlin
authors.initials: J
authors.lastname: Pan
authors.forename: Qingshun
authors.initials: Q
authors.lastname: Zhao
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002397:Catfishes / Q000196:embryology / Q000235:genetics* / Q000378:metabolism; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D004625:Embryo, Nonmammalian / Q000378:metabolism; D004720:Endonucleases / Q000378:metabolism*; D005260:Female; D029941:Fish Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D055786:Gene Knockout Techniques; D018390:Gene Targeting; D040582:Inheritance Patterns; D008297:Male; D008969:Molecular Sequence Data; D055435:Myostatin / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D010802:Phylogeny
chemical_list: D018076:DNA, Complementary; D029941:Fish Proteins; D055435:Myostatin; D004720:Endonucleases