提高哺乳动物细胞中基因编码 Ca2+ 生物传感器响应性的自动筛选平台

IF 3.5 Q2 CHEMISTRY, ANALYTICAL
Yufeng Zhao, Yi Shen, Teodor Veres and Robert E. Campbell
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引用次数: 0

摘要

基于基因编码的荧光蛋白(FP)生物传感器是对动态细胞活动进行成像的强大工具。定向进化是开发基于 FP 的增强型生物传感器的高效方法,但筛选过程费时费力。利用电刺激方法进行基于哺乳动物细胞的筛选已成功地准确筛选出用于成像神经元活动的生物传感器变体。我们介绍了一种利用荧光显微镜和液体分配器的自动化哺乳动物细胞筛选平台,以筛选生物传感器对化学刺激的反应性。我们展示了该平台在改善 Ca2+ 红色荧光生物传感器 K-GECO 的响应方面的有效性,该传感器可用于检测组胺诱导的 Ca2+ 浓度变化。这种方法适用于任何能对药物治疗或其他外源化学刺激做出反应的基于 FP 的生物传感器,从而简化了为不同生物环境中的特定应用开发生物传感器的工作。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

An automated screening platform for improving the responsiveness of genetically encoded Ca2+ biosensors in mammalian cells†

An automated screening platform for improving the responsiveness of genetically encoded Ca2+ biosensors in mammalian cells†

Genetically-encoded, fluorescent protein (FP)-based biosensors are powerful tools for imaging dynamic cellular activities. Directed evolution is a highly effective method for developing enhanced versions of FP-based biosensors, but the screening process is laborious and time-consuming. Mammalian cell-based screening with electrical stimulation methods has been successful in accurately selecting variants of biosensors for imaging neuronal activities. We introduce an automated mammalian cell screening platform utilizing a fluorescence microscope and a liquid dispenser to enable the screening of biosensor responsiveness to chemical stimulation. We demonstrated the effectiveness of this platform in improving the response of a red fluorescent biosensor for Ca2+, K-GECO, for detection of histamine-induced changes in Ca2+ concentration. This method should be applicable to any FP-based biosensor that responds to pharmacological treatment or other exogenous chemical stimulation, simplifying efforts to develop biosensors tailored for specific applications in diverse biological contexts.

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CiteScore
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