Developmental Biology Unit, European Molecular Biology Laboratory, Heidelberg 69117, Germany.
Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg 69117, Germany.
Proc Natl Acad Sci U S A. 2023 Jan 3;120(1):e2215958120. doi: 10.1073/pnas.2215958120. Epub 2022 Dec 27.
The cnidarian has developed into a powerful model system to study the mechanisms underlying animal development, regeneration, and evolution. However, despite the significant progress in the molecular and genetic approaches in this sea anemone, endogenous protein tagging is still challenging. Here, we report a robust method for knock in for using CRISPR/Cas9. As an outcome, we generate endogenously tagged proteins that label core molecular components of several cellular apparatus, including the nuclear envelope, cytoskeleton, cell adhesion, endoplasmic reticulum, cell trafficking, and extracellular matrix. Using live imaging, we monitor the dynamics of vesicular trafficking and endoplasmic reticulum in embryos, as well as cell contractility during the peristaltic wave of a primary polyp. This advancement in gene editing expands the molecular tool kit of and enables experimental avenues to interrogate the cell biology of cnidarians.
刺胞动物已经发展成为一个强大的模式系统，用于研究动物发育、再生和进化的机制。然而，尽管在这种海葵的分子和遗传方法上取得了重大进展，但内源性蛋白质标记仍然具有挑战性。在这里，我们报告了一种使用 CRISPR/Cas9 进行基因敲入的稳健方法。作为结果，我们生成了内源性标记的蛋白质，标记了几个细胞器官的核心分子成分，包括核膜、细胞骨架、细胞黏附、内质网、细胞运输和细胞外基质。通过活体成像，我们监测了胚胎中囊泡运输和内质网的动态，以及原发性息肉蠕动波期间的细胞收缩性。这项基因编辑的进展扩展了 的分子工具包，并为实验途径提供了研究刺胞动物细胞生物学的机会。
