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Published on: January 20, 2018
Generation of myostatin B knockout yellow catfish (Tachysurus fulvidraco) using transcription activator-like effector
Zhangji Dong1, Jiachun Ge, Zhiqiang Xu
11 MOE Key Laboratory of Model Animal for Disease Study, Model Animal Research Center, Nanjing University , Nanjing, China .
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.
Insights
Researchers identified and knocked out the second myostatin gene (mstnb) in yellow catfish. This study provides insights into the function of myostatin genes in fish growth and muscle development.
Area of Science:
- Genetics
- Developmental Biology
- Aquaculture
Background:
- Myostatin (Mstn) inhibits muscle growth in mammals, leading to a double-muscle phenotype when mutated.
- Myostatin's role in fish is less understood due to a lack of naturally occurring null alleles.
- Previous work involved knocking out mstna in yellow catfish.
Purpose of the Study:
- To identify and characterize the second myostatin gene (mstnb) in yellow catfish.
- To generate mstnb knockout yellow catfish using gene-editing technology.
- To investigate the potential role of mstnb in fish muscle development.
Main Methods:
- Identification and sequencing of the mstnb gene.
- Gene knockout using transcription activator-like effector nucleases (TALEN) technology.
- Analysis of mutated mstnb alleles, including frameshift and nonframeshift mutations.
Main Results:
- The mstnb gene was identified, comprising three exons and encoding 373 amino acids.
- TALEN technology successfully generated mstnb knockout yellow catfish founders.
- Six mutated mstnb alleles were identified, including two predicted to result in a functional null protein.
Conclusions:
- This study successfully generated mstnb knockout yellow catfish, providing a valuable tool for future research.
- The findings contribute to understanding the conserved and divergent roles of myostatin genes in vertebrate muscle growth.
- The generated mutants will aid in elucidating the specific functions of mstnb in fish development and aquaculture.
