Biozentrum der Universität Basel, Basel, Switzerland.
Methods Mol Biol. 2022;2540:219-237. doi: 10.1007/978-1-0716-2541-5_10.
The direct manipulation of proteins by nanobodies and other protein binders has become an additional and valuable approach to investigate development and homeostasis in Drosophila. In contrast to other techniques, that indirectly interfere with proteins via their nucleic acids (CRISPR, RNAi, etc.), protein binders permit direct and acute protein manipulation. Since the first use of a nanobody in Drosophila a decade ago, many different applications exploiting protein binders have been introduced. Most of these applications use nanobodies against GFP to regulate GFP fusion proteins. In order to exert specific protein manipulations, protein binders are linked to domains that confer them precise biochemical functions. Here, we reflect on the use of tools based on protein binders in Drosophila. We describe their key features and provide an overview of the available reagents. Finally, we briefly explore the future avenues that protein binders might open up and thus further contribute to better understand development and homeostasis of multicellular organisms.
纳米抗体和其他蛋白质结合物对蛋白质的直接操作已成为研究果蝇发育和动态平衡的另一种有价值的方法。与通过核酸（CRISPR、RNAi 等）间接干扰蛋白质的其他技术不同，蛋白质结合物允许直接和急性蛋白质操作。自十年前在果蝇中首次使用纳米抗体以来，已经引入了许多利用蛋白质结合物的不同应用。这些应用中的大多数都使用针对 GFP 的纳米抗体来调节 GFP 融合蛋白。为了进行特定的蛋白质操作，将蛋白质结合物与赋予它们精确生化功能的结构域相连。在这里，我们反思了在果蝇中使用基于蛋白质结合物的工具。我们描述了它们的关键特征，并提供了可用试剂的概述。最后，我们简要探讨了蛋白质结合物可能开辟的未来途径，从而为更好地理解多细胞生物的发育和动态平衡做出进一步贡献。
