Department of Pathology, Johns Hopkins Medical Institutions, Baltimore, MD, USA.
Weill Cornell Medicine, New York, NY, USA.
Methods Mol Biol. 2024;2800:189-202. doi: 10.1007/978-1-0716-3834-7_13.
Understanding how signaling networks are regulated offers valuable insights into how cells and organisms react to internal and external stimuli and is crucial for developing novel strategies to treat diseases. To achieve this, it is necessary to delineate the intricate interactions between the nodes in the network, which can be accomplished by measuring the activities of individual nodes under perturbation conditions. To facilitate this, we have recently developed a biosensor barcoding technique that enables massively multiplexed tracking of numerous signaling activities in live cells using genetically encoded fluorescent biosensors. In this chapter, we detail how we employed this method to reconstruct the EGFR signaling network by systematically monitoring the activities of individual nodes under perturbations.
了解信号转导网络如何被调控，能够为我们深入理解细胞和生物体如何对外界和内部刺激做出反应提供有价值的线索，并且对于开发新的疾病治疗策略至关重要。要实现这一目标，需要描绘出网络中节点之间复杂的相互作用，这可以通过在扰动条件下测量单个节点的活性来完成。为了实现这一目标，我们最近开发了一种生物传感器条码技术，该技术可以使用遗传编码的荧光生物传感器在活细胞中大规模地、多重地跟踪许多信号活性。在本章中，我们详细介绍了如何通过系统地监测扰动下单个节点的活性，来利用该方法重建 EGFR 信号转导网络。
