文献类型：期刊文献

英文题名：Monitoring cellular redox state under hypoxia using a fluorescent sensor based on eel fluorescent protein

作者：Hu, Hanyang[1,3];Wang, Aoxue[1,3];Huang, Li[1,3];Zou, Yejun[1,3];Gu, Yanfang[1,3];Chen, Xianjun[1,3];Zhao, Yuzheng[1,3];Yang, Yi[1,2]

机构：[1]East China Univ Sci & Technol, Shanghai Collaborat Innovat Ctr Biomfg Technol, State Key Lab Bioreactor Engn, Synthet Biol & Biotechnol Lab, 130 Mei Long Rd, Shanghai 200237, Peoples R China;[2]Chinese Acad Sci, Shanghai Inst Biol Sci, CAS Ctr Excellence Brain Sci, Optogenet & Synthet Biol Interdisciplinary Res Ct, Shanghai 200031, Peoples R China;[3]East China Univ Sci & Technol, Sch Pharm, Shanghai Key Lab New Drug Design, 130 Mei Long Rd, Shanghai 200237, Peoples R China

年份：2018

卷号：120

起止页码：255

外文期刊名：FREE RADICAL BIOLOGY AND MEDICINE

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000431771500025)】；

基金：We thank S. James Remington for the roGFP1 vector. This research was supported by National Key Research and Development Program of China (2017YFA050400, 2017YFC0906900), NSFC (31722033, 91649123, 31671484, 31225008, and 31470833), the Shanghai Science and Technology Commission (14XD1401400, 16430723100, and 15YF1402600), Young Elite Scientists Sponsorship Program by Cast (to Y. Zhao), Shanghai Young Top-notch Talent (to Y. Zhao), the State Key Laboratory of Bioreactor Engineering (to Y. Y.), and the Fundamental Research Funds for the Central Universities (to Y. Y. and Y. Zhao).

语种：英文

外文关键词：Eel fluorescent protein UnaG; Redox state; Real-time monitoring; RoUnaG; Oxygen-independent; Hypoxia

摘要：Genetically encoded fluorescent sensors are widely used to visualize secondary messengers, metabolites and dynamic events in living cells. However, almost all of these sensors are based on Aequorea GFPs or GFP-like proteins, which do not correctly maturate and fluoresce under hypoxia or anoxic conditions, greatly limiting their application in biomedical research. Herein, we provide a novel strategy for design of sensors and report a series of thiol redox-sensitive sensor based on a recently discovered oxygen-independent fluorescent protein UnaG from Japanese eel. These redox sensors have large dynamic range, rapid responsiveness, a flexible "switch", and pH-independence, are particularly compatible with hypoxia conditions, and therefore represent a substantial improvement for live-cell redox measurement. We further demonstrated the versatility of these redox sensors, by simultaneously monitoring redox changes and hypoxia state in living cells, thereby proving its capability as a powerful and flexible tool for indexing multidimensional metabolism data in the context of physiological stressors and pathological states. These redox sensors are not only the first case of UnaG-based functional sensors, but also the first case of functional sensors based on non GFP-like proteins. Based on this strategy, more oxygen-independent biosensors could be developed, hence, provide new opportunities for bioimaging.