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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. © 2024 The Author(s). Advanced Science published by Wiley‐VCH GmbH.

Citation

Yuanfa Yao, Xiayan Lou, Luhong Jin, Weiyun Sun, Jingfang Liu, Yunyue Chen, Sunying Cheng, Tengjiao Zhao, Shuwei Ke, Luhao Zhang, Yingke Xu, Lian He, Hanbing Li. Optogenetic Strategies for Optimizing the Performance of Phospholipids Biosensors. Advanced science (Weinheim, Baden-Wurttemberg, Germany). 2024 Sep;11(36):e2403026

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PMID: 39073033

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