Rapid and sustained degradation of the essential centrosome protein CEP192 in live mice using the AID2 system

IF 12.5 1区综合性期刊 Q1 MULTIDISCIPLINARY SCIENCES

Science Advances Pub Date : 2025-02-28 DOI:10.1126/sciadv.adq2339

Valentina C. Sladky, Margaret A. Strong, Daniel Tapias-Gomez, Cayla E. Jewett, Chelsea G. Drown, Phillip M. Scott, Andrew J. Holland

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Abstract

Studying essential genes required for dynamic processes in live mice is challenging as genetic perturbations are irreversible and limited by slow protein depletion kinetics. The auxin-inducible degron (AID) system is a powerful tool for analyzing inducible protein loss in vitro, but it is toxic to mice. Here, we use an optimized second-generation AID system to achieve the conditional and reversible loss of the essential centrosomal protein CEP192 in live mice. We show that the auxin derivative 5-phenyl-indole-3-acetic acid is well tolerated over 2 weeks and drives near-complete CEP192 degradation in less than 1 hour in vivo. CEP192 loss did not affect centriole duplication but decreased γ-tubulin recruitment to centrosomes impairing mitotic spindle assembly. Sustained CEP192 loss in vivo led to cell division failure and cell death in proliferative tissues. Thus, the second-generation AID system is well suited for rapid and/or sustained protein depletion in live mice to study essential functions in vivo.

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使用AID2系统快速和持续地降解活小鼠必需中心体蛋白CEP192

在活体小鼠体内研究动态过程所需的重要基因具有挑战性，因为基因扰动是不可逆的，而且受限于缓慢的蛋白质损耗动力学。辅助因子诱导去势（AID）系统是分析体外诱导性蛋白质损失的强大工具，但它对小鼠有毒。在这里，我们使用优化的第二代 AID 系统在活体小鼠体内实现了重要中心粒蛋白 CEP192 的条件性和可逆性缺失。我们的研究表明，辅助素衍生物 5-苯基-吲哚-3-乙酸在 2 周内具有良好的耐受性，并能在体内不到 1 小时的时间内促使 CEP192 几乎完全降解。CEP192 的缺失不会影响中心粒的复制，但会减少 γ-微管蛋白对中心体的招募，从而损害有丝分裂纺锤体的组装。在体内持续缺失 CEP192 会导致细胞分裂失败和增殖组织中的细胞死亡。因此，第二代 AID 系统非常适合在活体小鼠体内快速和/或持续地消耗蛋白质，以研究体内的基本功能。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Science Advances 综合性期刊-综合性期刊

CiteScore

21.40

自引率

1.50%

发文量

1937

审稿时长

29 weeks

期刊介绍： Science Advances, an open-access journal by AAAS, publishes impactful research in diverse scientific areas. It aims for fair, fast, and expert peer review, providing freely accessible research to readers. Led by distinguished scientists, the journal supports AAAS's mission by extending Science magazine's capacity to identify and promote significant advances. Evolving digital publishing technologies play a crucial role in advancing AAAS's global mission for science communication and benefitting humankind.