ANRI, Tokyo, Japan.
Methods Mol Biol. 2021;2312:277-285. doi: 10.1007/978-1-0716-1441-9_16.
There are increasing evidence and growing interest in the relationship between protein aggregates/phase separation and various human diseases, especially neurodegenerative diseases. However, we do not entirely comprehend how aggregates generate or the clearance network of chaperones, proteasomes, ubiquitin ligases, and other factors interact with aggregates. Here, we describe chemically controllable systems compose with a genetically engineered cell and a small drug that enables us to rapidly induce protein aggregates' formation by withdrawing the small molecule. This trigger does not activate global stress responses induced by stimuli, such as proteasome inhibitors or heat shock. This method can produce aggregates in a specific compartment and diverse experimental systems, including live animals.
越来越多的证据和研究兴趣表明，蛋白质聚集/相分离与各种人类疾病（尤其是神经退行性疾病）之间存在关联。然而，我们并不完全理解聚集物是如何产生的，也不了解伴侣蛋白、蛋白酶体、泛素连接酶和其他因子的清除网络如何与聚集物相互作用。在这里，我们描述了一个由经过基因工程改造的细胞和一种小分子药物组成的化学控制体系，该体系可以通过去除小分子来快速诱导蛋白质聚集物的形成。这种触发不会激活由蛋白酶体抑制剂或热休克等刺激引起的全局应激反应。该方法可以在特定的隔间和多种实验系统中产生聚集物，包括活体动物。
