A Method for the Acute and Rapid Degradation of Endogenous Proteins.,Cell

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A Method for the Acute and Rapid Degradation of Endogenous Proteins.
Cell ( IF 45.5 ) Pub Date : 2017-Dec-14 , DOI: 10.1016/j.cell.2017.10.033
Dean Clift ₁ , William A McEwan ₁ , Larisa I Labzin ₁ , Vera Konieczny ₂ , Binyam Mogessie ₂ , Leo C James ₁ , Melina Schuh ₃

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Methods for the targeted disruption of protein function have revolutionized science and greatly expedited the systematic characterization of genes. Two main approaches are currently used to disrupt protein function: DNA knockout and RNA interference, which act at the genome and mRNA level, respectively. A method that directly alters endogenous protein levels is currently not available. Here, we present Trim-Away, a technique to degrade endogenous proteins acutely in mammalian cells without prior modification of the genome or mRNA. Trim-Away harnesses the cellular protein degradation machinery to remove unmodified native proteins within minutes of application. This rapidity minimizes the risk that phenotypes are compensated and that secondary, non-specific defects accumulate over time. Because Trim-Away utilizes antibodies, it can be applied to a wide range of target proteins using off-the-shelf reagents. Trim-Away allows the study of protein function in diverse cell types, including non-dividing primary cells where genome- and RNA-targeting methods are limited.

中文翻译：

一种快速降解内源蛋白质的方法。

靶向破坏蛋白质功能的方法彻底改变了科学，并极大地加快了基因的系统表征。目前破坏蛋白质功能的主要方法有两种：DNA 敲除和 RNA 干扰，分别作用于基因组和 mRNA 水平。目前还没有直接改变内源蛋白质水平的方法。在这里，我们提出了 Trim-Away，一种在哺乳动物细胞中急剧降解内源蛋白的技术，无需事先修改基因组或 mRNA。Trim-Away 利用细胞蛋白质降解机制在应用后几分钟内去除未修饰的天然蛋白质。这种速度最大限度地降低了表型被补偿以及继发性非特异性缺陷随时间累积的风险。由于 Trim-Away 利用抗体，因此可以使用现成试剂将其应用于多种目标蛋白。Trim-Away 允许研究不同细胞类型中的蛋白质功能，包括基因组和 RNA 靶向方法有限的非分裂原代细胞。

更新日期：2017-11-19

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