Generation of fast growth Nile tilapia (Oreochromis niloticus) by myostatin gene mutation
Generation of fast growth Nile tilapia (Oreochromis niloticus) by myostatin gene mutation
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通过肌生长抑制素基因突变培育快速生长的尼罗罗非鱼（Oreochromis niloticus）
DOI：
10.1016/j.aquaculture.2022.738762
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发表时间：
2022-08
期刊：
影响因子：
4.5
通讯作者：
Zhou Linyan
中科院分区：
文献类型：
--
作者：
Wu You;Wu Tengfei;Yang Lanying;Su Yun;Zhao Chenhua;Li Lu;Cai Jing;Dai Xiangyan;Wang Deshou;Zhou Linyan
罗非鱼是水产养殖中产量第二高的品种，仅次于鲤鱼，在超过 100 个国家广泛种植。肌肉生长抑制素（MSTN）已被证明是骨骼肌生长的负调节因子。 MSTN 基因突变导致脊椎动物的体型和肌肉质量显着增加，且大多具有物种特异性效应。为了培育具有快速生长特性的尼罗罗非鱼（Oreochromis niloticus）新品系，对mstn基因的分子表征、基因表达和功能研究进行了广泛的研究。系统发育和同线性分析表明，mstnb 可能是哺乳动物MSTN 基因的直系同源基因，而mstna 则不然。 qRT-PCR 分析的表达模式表明，mstnb 在肌肉中丰富，而 mstna 主要在大脑中表达。荧光原位杂交（FISH）分析还表明mstnb仅在白色骨骼肌纤维的基底膜中表达，进一步提示mstnb基因在罗非鱼肌肉生长中的可能作用。为了验证mstnb在骨骼肌生长中的作用，我们利用CRISPR/Cas9基因编辑技术获得了mstnb −/− 突变体。与野生型（WT）鱼相比，mstnb -/− 突变体从孵化后 5 个月（mah）起，表现出典型的双肌表型，肌肉质量增加，在头部和背鳍之间突出。形态学观察显示，7 mah 时白色肌纤维过度增殖。 mstnb−/−鱼的平均体重（21.67±8.26g）、体高（4.46±0.65cm）和体宽（2.36±0.53cm）分别比mstnb+/+鱼（体重14.50±8.60）高49.45%、32.74%和37.21%。 g；5mAh时，体高3.36±0.45cm；体宽1.72±0.18cm。结果表明，在5 mah的实验室条件下，mstnb −/−鱼的生长性能参数，包括增重率（1.99倍）、条件因子（1.77倍）和特定生长率（1.23倍）均显着高于mstnb +/+鱼。此外，在野生自然环境下90天的饲养试验中，mstnb−/−尼罗罗非鱼的WGR、CF、SGR和FE参数显着高于mstnb+/+尼罗罗非鱼和GIFT罗非鱼。有趣的是，在 7 mah 时，随着 mstn −/− XY 鱼骨骼肌生长性能的提高，发现了对肌肉生长的性别二态性影响，但 mstn −/− XX 鱼却没有。综上所述，本研究首次证明mstn基因突变可能促进尼罗罗非鱼快速生长雄性品系的产生。 • 通过CRISPR/cas9 技术在尼罗罗非鱼中产生纯合mstnb 突变系。 • Mstnb 缺乏导致罗非鱼的生长促进偏向雄性。 • mstnb −/− 罗非鱼的WGR、CF、SGR 和FE 高于mstnb +/+ 和GIFT 罗非鱼。
Tilapia is the second most prolific species grown in aquaculture after carp, and is widely grown in >100 countries. Myostatin (MSTN) has been proved to be a negative regulator of skeletal muscle growth. Mutation of MSTN gene resulted in significant increase in both body size and muscle mass in vertebrates, mostly with a species-specific effect. To generate a new strain in Nile tilapia ( Oreochromis niloticus ) with fast growth traits, the molecular characterization, gene expression and functional studies of mstn gene were extensively investigated. Phylogenetic and synteny analyses indicated that mstnb , but not mstna , might be the orthologous gene of mammalian MSTN gene. Expression pattern analyzed by qRT-PCR demonstrated that mstnb is abundant in muscle, while mstna is dominantly expressed in the brain. Fluorescence in situ hybridization ( FISH ) analysis also demonstrated that mstnb is exclusively expressed in the basement membrane of white skeletal muscle fibers, further suggesting the possible role of mstnb gene in tilapia muscle growth. To verify the roles of mstnb in skeletal muscle growth, we obtain the mstnb −/− mutant using CRISPR/Cas9 gene editing technology. Compared with the wild type (WT) fish, the mstnb −/− mutants showed a typical double-muscle phenotype with increased muscle mass, sticking out between head and dorsal fin, from 5 months after hatching (mah). Morphological observation revealed an excessive proliferation of white muscle fibers at 7 mah. The average body weights (21.67 ± 8.26 g), body heights (4.46 ± 0.65 cm) and body widths (2.36 ± 0.53 cm) of the mstnb −/− fish were 49.45%, 32.74% and 37.21% higher than those of the mstnb +/+ fish (body weight, 14.50 ± 8.60 g; body height, 3.36 ± 0.45 cm; body width, 1.72 ± 0.18 cm) at 5 mah. The results showed that the growth performance parameters of the mstnb −/− fish, including weight gain rate (1.99 times), condition factor (1.77 times) and specific growth rate (1.23 times), were significantly higher than the mstnb +/+ fish under laboratory conditions at 5 mah. In addition, the parameters of WGR, CF, SGR and FE were significantly increased in mstnb −/− Nile tilapia than those of mstnb +/+ Nile tilapia and GIFT tilapia for a 90-day feeding trial under the wild natural environment. Interestingly, sexually dimorphic effects on muscle growth have been found with an increase of growth performance of skeletal muscle in mstn −/− XY fish, but not mstn −/− XX fish at 7 mah. Taken together, this study, for the first time, proved that mstn gene mutation might promote the generation of new fast-growing male strain of Nile tilapia. • Homozygous mstnb mutant line was generated by CRISPR/cas9 technology in Nile tilapia. • Mstnb deficiency resulted in male biased growth enhancement in tilapia. • The mstnb −/− tilapia show a higher WGR, CF, SGR and FE than mstnb +/+ and GIFT tilapia.
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DOI：
10.29271/jcpsp.2018.06.419
发表时间：
2018-05
期刊：
Journal of the College of Physicians and Surgeons--Pakistan : JCPSP
影响因子：
--
作者：
Hamd Binte Shahab Syed;Muhammad Yunus Khan
通讯作者：
Hamd Binte Shahab Syed;Muhammad Yunus Khan
使用 CRISPR/Cas9 系统有效敲除山羊基因
影响因子：
3.7
作者：
Ni W;Qiao J;Hu S;Zhao X;Regouski M;Yang M;Polejaeva IA;Chen C
通讯作者：
Chen C
影响因子：
4.8
作者：
Kim, Gap-Don;Lee, Jeong Hyo;Park, Tae Sub
通讯作者：
Park, Tae Sub
影响因子：
--
作者：
Eckhardt, Katrin;Troeger, Juliane;Wenger, Roland H.
通讯作者：
Wenger, Roland H.
影响因子：
--
作者：
Indranath Ghosal;S. Chakraborty
通讯作者：
Indranath Ghosal;S. Chakraborty