论文标题
ARF-AID系统：保留内源性蛋白水平并促进迅速诱导蛋白降解的方法
The ARF-AID system: Methods that preserve endogenous protein levels and facilitate rapidly inducible protein degradation
论文作者
论文摘要
ARF-AID（生长素反应因子 - 铝诱导脱哥伦斯）系统是一种重新设计的生长素诱导蛋白降解系统。诱导型脱哥伦斯系统被广泛用于细胞系和生物中感兴趣的蛋白质的特定和快速消耗。诱导降解的一个优点是，所研究的生物系统在扰动之前保持完整和功能。此功能需要维持内源性水平。然而，用辅助的基因内源性标记会导致慢性，与生长素无关的蛋白酶体介导的降解。在重新设计的ARF-AID系统中，ARF-PB1结构域的附加表达可防止艾滋病标记蛋白的长期降解，同时保留标记蛋白的快速降解。在这里，我们描述了工程人类细胞系实施ARF-AID系统的特定蛋白质降解的方案。这些方法是适应性的，可以从细胞系扩展到生物。
The ARF-AID (Auxin Response Factor-Auxin Inducible Degron) system is a re-engineered auxin-inducible protein degradation system. Inducible degron systems are widely used to specifically and rapidly deplete proteins of interest in cell lines and organisms. An advantage of inducible degradation is that the biological system under study remains intact and functional until perturbation. This feature necessitates that the endogenous levels of the protein are maintained. However, endogenous tagging of genes with AID can result in chronic, auxin-independent proteasome-mediated degradation. The additional expression of the ARF-PB1 domain in the re-engineered ARF-AID system prevents chronic degradation of AID-tagged proteins while preserving rapid degradation of tagged proteins. Here we describe the protocol for engineering human cell lines to implement the ARF-AID system for specific and inducible protein degradation. These methods are adaptable and can be extended from cell lines to organisms.
