Faculty of Biology, Moscow State University, Moscow, Russia; Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Moscow 117997, Russia.
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Moscow 117997, Russia; Pirogov Russian National Research Medical University, Moscow 117997, Russia.
Free Radic Biol Med. 2018 Nov 20;128:23-39. doi: 10.1016/j.freeradbiomed.2018.04.004. Epub 2018 Apr 6.
A wide variety of genetically encoded fluorescent biosensors are available to date. Some of them have already contributed significantly to our understanding of biological processes occurring at cellular and organismal levels. Using such an approach, outstanding success has been achieved in the field of redox biology. The probes allowed researchers to observe, for the first time, the dynamics of important redox parameters in vivo during embryogenesis, aging, the inflammatory response, the pathogenesis of various diseases, and many other processes. Given the differences in the readout and spectra of the probes, they can be used in multiparameter imaging in which several processes are monitored simultaneously in the cell. Intracellular processes form an extensive network of interactions. For example, redox changes are often accompanied by changes in many other biochemical reactions related to cellular metabolism and signaling. Therefore, multiparameter imaging can provide important information concerning the temporal and spatial relationship of various signaling and metabolic processes. In this review, we will describe the main types of genetically encoded biosensors, the most frequently used readout, and their use in multiplexed imaging mode.
目前已有多种基因编码荧光生物传感器可供选择。其中一些已经为我们理解细胞和生物水平上发生的生物学过程做出了重大贡献。通过这种方法,在氧化还原生物学领域取得了卓越的成功。这些探针使研究人员能够首次在胚胎发生、衰老、炎症反应、各种疾病的发病机制以及许多其他过程中观察到体内重要氧化还原参数的动态变化。鉴于探针的读出和光谱存在差异,它们可用于多参数成像,其中可以同时在细胞中监测多个过程。细胞内过程形成了广泛的相互作用网络。例如,氧化还原变化通常伴随着与细胞代谢和信号转导相关的许多其他生化反应的变化。因此,多参数成像可以提供有关各种信号转导和代谢过程的时空关系的重要信息。在这篇综述中,我们将描述主要类型的基因编码生物传感器、最常使用的读出方法及其在多路复用成像模式中的应用。