title: TALENs-mediated gene disruption of myostatin produces a larger phenotype of medaka with an apparently compromised immune system.
abstract: Although myostatin, a suppressor of skeletal muscle development and growth, has been well studied in mammals, its function in fish remains unclear. In this study, we used a popular genome editing tool with high efficiency and target specificity (TALENs; transcription activator-like effector nucleases) to mutate the genome sequence of myostatin (MSTN) in medaka (Oryzias latipes). After the TALEN pair targeting OlMyostatin was injected into fertilized medaka eggs, mutant G0 fish carrying different TALENs-induced frameshifts in the OlMSTN coding sequence were mated together in order to transmit the mutant sequences to the F1 generation. Two F1 mutants with frameshifted myostatin alleles were then mated to produce the F2 generation, and these F2 OlMSTN null (MSTN(-/-)) medaka were evaluated for growth performance. The F2 fish showed significantly increased body length and weight compared to the wild type fish at the juvenile and post-juvenile stages. At the post-juvenile stage, the average body weight of the MSTN(-/-) medaka was ∼25% greater than the wild type. However, we also found that when the F3 generation were challenged with red spotted grouper nervous necrosis virus (RGNNV), the expression levels of the interferon-stimulated genes were lower than in the wild type, and the virus copy number was maintained at a high level. We therefore conclude that although the MSTN(-/-) medaka had a larger phenotype, their immune system appeared to be at least partially suppressed or undeveloped.
authors.affiliation: Institute of Biotechnology, College of Bioscience and Biotechnology, National Cheng Kung University, Tainan 701, Taiwan.
authors.forename: Yi-An
authors.initials: YA
authors.lastname: Chiang
authors.affiliation: Division of Applied Bioscience, Graduate School of Agriculture, Kyoto University, Kyoto 606-8502, Japan.
authors.forename: Masato
authors.initials: M
authors.lastname: Kinoshita
authors.affiliation: Institute of Biotechnology, College of Bioscience and Biotechnology, National Cheng Kung University, Tainan 701, Taiwan.
authors.forename: Shun
authors.initials: S
authors.lastname: Maekawa
authors.affiliation: Institute of Biotechnology, College of Bioscience and Biotechnology, National Cheng Kung University, Tainan 701, Taiwan.
authors.forename: Amod
authors.initials: A
authors.lastname: Kulkarni
authors.affiliation: Institute of Bioinformatics and Biosignal Transduction, National Cheng Kung University, Tainan City 701, Taiwan.
authors.forename: Chu-Fang
authors.initials: CF
authors.lastname: Lo
authors.affiliation: National Research Institute of Fisheries and Enhancement of Inland Sea, Fisheries Research Agency, Kagawa 761-0111, Japan.
authors.forename: Yasutoshi
authors.initials: Y
authors.lastname: Yoshiura
authors.affiliation: Institute of Biotechnology, College of Bioscience and Biotechnology, National Cheng Kung University, Tainan 701, Taiwan. Electronic address: wanghc@mail.ncku.edu.tw.
authors.forename: Han-Ching
authors.initials: HC
authors.lastname: Wang
authors.affiliation: Institute of Biotechnology, College of Bioscience and Biotechnology, National Cheng Kung University, Tainan 701, Taiwan. Electronic address: takashi.aoki65@gmail.com.
authors.forename: Takashi
authors.initials: T
authors.lastname: Aoki
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D049628:Body Size; D003851:Deoxyribonucleases / Q000235:genetics; D005260:Female; D005393:Fish Diseases / Q000235:genetics / Q000276:immunology / Q000821:virology; D029941:Fish Proteins / Q000235:genetics* / Q000276:immunology*; D007372:Interferons / Q000276:immunology; D008297:Male; D055435:Myostatin / Q000235:genetics* / Q000276:immunology*; D030381:Nodaviridae; D009990:Oryzias / Q000235:genetics / Q000254:growth & development / Q000276:immunology / Q000821:virology; D010641:Phenotype; D012327:RNA Virus Infections / Q000235:genetics / Q000276:immunology / Q000662:veterinary / Q000821:virology
chemical_list: D029941:Fish Proteins; D055435:Myostatin; D007372:Interferons; D003851:Deoxyribonucleases
keywords: Larger phenotype; Medaka; Myostatin; Transcription activator-like effector nucleases (TALENs); Weaker immune system