ARF-AID系统：保持内源性蛋白水平并促进快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
摘要：
ARF-AID（AuxinResponseFactor-AuxinInducible Degron）系统是一个重组的生长素诱导蛋白降解系统。诱导型降解决定子系统广泛用于特异性地和快速地耗尽细胞系和生物体中的感兴趣的蛋白质。诱导降解的一个优点是，所研究的生物系统保持完整和功能，直到扰动。这一特征需要维持蛋白质的内源性水平。然而，内源性标记的基因与AID可导致慢性，生长素不依赖性蛋白酶体介导的降解。在重新设计的ARF-AID系统中ARF-PB 1结构域的额外表达防止了AID标记蛋白的慢性降解，同时保持了标记蛋白的快速降解。在这里，我们描述了工程人类细胞系的协议，实现特异性和诱导蛋白质降解的ARF-AID系统。这些方法适应性强，可以从细胞系扩展到生物体。
英文摘要：
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.
机构、总结、中文摘要均由AI分析而来，可能存在错误，请以论文正文为准。
