Department of Anatomy, Korea University College of Medicine, Seoul, Republic of Korea.
BK21 Graduate Program, Department of Biomedical Sciences, Korea University College of Medicine, Seoul, Republic of Korea.
Nat Chem Biol. 2024 Mar;20(3):353-364. doi: 10.1038/s41589-023-01480-6. Epub 2023 Nov 16.
Proteases function as pivotal molecular switches, initiating numerous biological events. Notably, potyviral protease, derived from plant viruses, has emerged as a trusted proteolytic switch in synthetic biological circuits. To harness their capabilities, we have developed a single-component photocleavable switch, termed LAUNCHER (Light-Assisted UNcaging switCH for Endoproteolytic Release), by employing a circularly permutated tobacco etch virus protease and a blue-light-gated substrate, which are connected by fine-tuned intermodular linkers. As a single-component system, LAUNCHER exhibits a superior signal-to-noise ratio compared with multi-component systems, enabling precise and user-controllable release of payloads. This characteristic renders LAUNCHER highly suitable for diverse cellular applications, including transgene expression, tailored subcellular translocation and optochemogenetics. Additionally, the plug-and-play integration of LAUNCHER into existing synthetic circuits facilitates the enhancement of circuit performance. The demonstrated efficacy of LAUNCHER in improving existing circuitry underscores its significant potential for expanding its utilization in various applications.
蛋白酶作为关键的分子开关，启动着众多的生物学事件。值得注意的是，来源于植物病毒的 Potyviral 蛋白酶已经成为合成生物学回路中可靠的蛋白水解开关。为了利用它们的功能，我们通过使用环状排列的烟草蚀纹病毒蛋白酶和蓝光门控底物，以及通过精细调节的模块间连接体，开发了一种称为 LAUNCHER（光辅助无笼开关用于内切蛋白酶释放）的单组分光可裂解开关。作为一种单组分系统，LAUNCHER 与多组分系统相比具有更高的信噪比，能够实现精确和用户可控的有效载荷释放。这种特性使 LAUNCHER 非常适合各种细胞应用，包括转基因表达、定制的亚细胞转位和光遗传药理学。此外，LAUNCHER 可插入到现有合成回路中，从而可以方便地增强回路性能。LAUNCHER 在改进现有电路方面的功效证明了它在各种应用中扩展其使用的巨大潜力。
