TY - JOUR
T1 - TALENs-mediated gene disruption of myostatin produces a larger phenotype of medaka with an apparently compromised immune system
AU - Chiang, Yi An
AU - Kinoshita, Masato
AU - Maekawa, Shun
AU - Kulkarni, Amod
AU - Lo, Chu Fang
AU - Yoshiura, Yasutoshi
AU - Wang, Han Ching
AU - Aoki, Takashi
N1 - Funding Information:
This study was supported financially by the Ministry of Science and Technology ( MOST 103-2633-B-006-004 ). We warmly thank Paul Barlow, National Cheng Kung University, for his helpful criticism of the manuscript.
Publisher Copyright:
© 2015 Elsevier Ltd.
PY - 2016/1/1
Y1 - 2016/1/1
N2 - 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.
AB - 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.
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U2 - 10.1016/j.fsi.2015.11.016
DO - 10.1016/j.fsi.2015.11.016
M3 - Article
C2 - 26578247
AN - SCOPUS:84949654025
VL - 48
SP - 212
EP - 220
JO - Fish and Shellfish Immunology
JF - Fish and Shellfish Immunology
SN - 1050-4648
ER -