Tianjin Key Laboratory of Animal and Plant Resistance, College of Life Science, Tianjin Normal University, Tianjin 300387, People's Republic of China.
J Exp Biol. 2022 Mar 15;225(6). doi: 10.1242/jeb.243702. Epub 2022 Mar 14.
CRISPR/Cas9 technology has been applied to many arthropods. However, application of this technology to crustaceans remains limited because of the unique characteristics of embryos. Our group has developed a microinjection system to introduce the CRISPR/Cas9 system into Neocaridina heteropoda embryos (one-cell stage). Using the developed method, we mutated the target gene Nh-scarlet (N. heteropoda scarlet), which functions in eye development and pigmentation. The results showed that both eye color and shape were altered in individuals in which Nh-scarlet was knocked out. Furthermore, this system was also successfully applied to another decapod crustacean, Eriocheir sinensis. DNA sequencing revealed that the zoeae with red eyes had an edited version of Es-scarlet. This study provides a stable microinjection method for freshwater crustaceans, and will contribute to functional genomics studies in various decapods.
CRISPR/Cas9 技术已被应用于许多节肢动物。然而，由于胚胎的独特特性，该技术在甲壳类动物中的应用仍然有限。我们小组开发了一种微注射系统，可将 CRISPR/Cas9 系统引入秀丽新小杆线虫胚胎（单细胞阶段）。使用所开发的方法，我们突变了目标基因 Nh-scarlet（秀丽新小杆线虫的 scarlet），该基因在眼睛发育和色素沉着中起作用。结果表明，Nh-scarlet 敲除的个体的眼睛颜色和形状都发生了改变。此外，该系统还成功应用于另一种十足甲壳类动物中华绒螯蟹。DNA 测序显示，具有红色眼睛的幼体具有编辑过的 Es-scarlet 版本。本研究为淡水甲壳类动物提供了一种稳定的微注射方法，将有助于各种十足甲壳类动物的功能基因组学研究。
