Generation of myostatin-knockout chickens mediated by D10A-Cas9 nickase
Generation of myostatin-knockout chickens mediated by D10A-Cas9 nickase
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DOI：
10.1096/fj.201903035r
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发表时间：
2020-02-25
期刊：
影响因子：
4.8
通讯作者：
Park, Tae Sub
中科院分区：
文献类型：
--
作者：
Kim, Gap-Don;Lee, Jeong Hyo;Park, Tae Sub
已经进行了许多研究来改善经济上重要的牲畜性状，如饲料效率和肌肉生长。基因组编辑技术代表了基础研究和农学生物技术发展的重大进步。成簇规则间隔短回文重复序列（CRISPR）/Cas9技术平台是用于工程化特定靶向基因座的强大工具。然而，脱靶效应的潜在发生，包括非预期靶标的切割，限制了Cas9介导的基因组编辑的实际应用。在这项研究中，为了最大限度地减少这种技术的脱靶效应，我们利用D10 A-Cas9切口酶通过原始生殖细胞产生肌肉生长抑制素敲除（MSTKO）鸡。D10 A-Cas9切口酶（Cas9 n）介导的MSTK 0鸡在胸部和腿部表现出显著更大的骨骼肌。肌肉生长抑制素缺失引起的骨骼肌肥大和增生程度因性别和肌肉类型而异。腹部脂肪沉积显着低于MSTKO鸡比野生型鸡。我们的研究结果表明，D10 A-Cas9技术平台可以促进精确和高效的靶向基因组工程，并可能扩大基因组编辑鸡在实际工业化和作为人类疾病动物模型中的应用范围。
Many studies have been conducted to improve economically important livestock traits such as feed efficiency and muscle growth. Genome editing technologies represent a major advancement for both basic research and agronomic biotechnology development. The clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 technical platform is a powerful tool used to engineer specific targeted loci. However, the potential occurrence of off-target effects, including the cleavage of unintended targets, limits the practical applications of Cas9-mediated genome editing. In this study, to minimize the off-target effects of this technology, we utilized D10A-Cas9 nickase to generate myostatin-knockout (MSTN KO) chickens via primordial germ cells. D10A-Cas9 nickase (Cas9n)-mediated MSTN KO chickens exhibited significantly larger skeletal muscles in the breast and leg. Degrees of skeletal muscle hypertrophy and hyperplasia induced by myostatin deletion differed by sex and muscle type. The abdominal fat deposition was dramatically lower in MSTN KO chickens than in wild-type chickens. Our results demonstrate that the D10A-Cas9 technical platform can facilitate precise and efficient targeted genome engineering and may broaden the range of applications for genome-edited chickens in practical industrialization and as animal models of human diseases.