Institute of Pharmacology, College of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou, 310014, China.
School of Information Science and Technology, Hangzhou Normal University, Hangzhou, Zhejiang, 311121, China.
Adv Sci (Weinh). 2024 Sep;11(36):e2403026. doi: 10.1002/advs.202403026. Epub 2024 Jul 29.
High-performance biosensors play a crucial role in elucidating the intricate spatiotemporal regulatory roles and dynamics of membrane phospholipids. However, enhancing the sensitivity and imaging performance remains a significant challenge. Here, optogenetic-based strategies are presented to optimize phospholipid biosensors. These strategies involves presequestering unbound biosensors in the cell nucleus and regulating their cytosolic levels with blue light to minimize background signal interference in phospholipid detection, particularly under conditions of high expression levels of biosensor. Furthermore, optically controlled phase separation and the SunTag system are employed to generate punctate probes for substrate detection, thereby amplifying biosensor signals and enhancing visualization of the detection process. These improved phospholipid biosensors hold great potential for enhancing the understanding of the spatiotemporal dynamics and regulatory roles of membrane lipids in live cells and the methodological insights in this study might be valuable for developing other high-performance biosensors.
高性能生物传感器在阐明膜磷脂的复杂时空调节作用和动态方面发挥着关键作用。然而，提高灵敏度和成像性能仍然是一个重大挑战。本文提出了基于光遗传学的策略来优化磷脂生物传感器。这些策略包括将未结合的生物传感器预隔离在细胞核内，并使用蓝光调节其细胞质水平，以最大限度地减少磷脂检测中的背景信号干扰，特别是在生物传感器表达水平较高的情况下。此外，还采用光控相分离和 SunTag 系统生成用于底物检测的点状探针，从而放大生物传感器信号并增强检测过程的可视化。这些改进后的磷脂生物传感器有望增强对活细胞中膜脂时空动态和调节作用的理解，本研究中的方法学见解对于开发其他高性能生物传感器可能具有重要价值。
