Key Laboratory of Mariculture, (Ocean University of China), Ministry of Education, Qingdao, 266003, China.
Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao, China.
Mar Biotechnol (NY). 2021 Apr;23(2):215-224. doi: 10.1007/s10126-020-10016-1. Epub 2021 Mar 14.
Pacific oyster (Crassostrea gigas) is one of the most widely cultivated shellfish species in the world. Because of its economic value and complex life cycle involving drastic changes from a free-swimming larva to a sessile juvenile, C. gigas has been used as a model for developmental, environmental, and aquaculture research. However, due to the lack of genetic tools for functional analysis, gene functions associated with biological or economic traits cannot be easily determined. Here, we reported a successful application of CRISPR/Cas9 technology for knockout of myosin essential light chain gene (CgMELC) in C. gigas. C. gigas embryos injected with sgRNAs/Cas9 contained extensive indel mutations at the target sites. The mutant larvae showed defective musculature and reduced motility. In addition, knockout of CgMELC disrupted the expression and patterning of myosin heavy chain positive myofibers in larvae. Together, these data indicate that CgMELC is involved in larval muscle contraction and myogenesis in oyster larvae.
太平洋牡蛎（Crassostrea gigas）是世界上养殖最广泛的贝类之一。由于其经济价值和涉及从自由游动的幼虫到固着幼体的剧烈变化的复杂生命周期，C. gigas 已被用作发育、环境和水产养殖研究的模型。然而，由于缺乏用于功能分析的遗传工具，与生物或经济性状相关的基因功能不能轻易确定。在这里，我们报告了 CRISPR/Cas9 技术在太平洋牡蛎肌球蛋白必需轻链基因（CgMELC）敲除中的成功应用。注射 sgRNAs/Cas9 的太平洋牡蛎胚胎在靶位点含有广泛的插入缺失突变。突变幼虫表现出肌肉缺陷和运动能力降低。此外，CgMELC 的敲除破坏了幼虫中肌球蛋白重链阳性肌纤维的表达和模式。总之，这些数据表明 CgMELC 参与了牡蛎幼虫的肌肉收缩和肌发生。
