Microbiology & Cell Biology
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Item A sensitive and specific genetically-encoded potassium ion biosensor for in vivo applications across the tree of life(Public Library of Science, 2022-09) Wu, Sheng-Yi; Wen, Yurong; Serre, Nelson B. C.; Heiede Laursen, Cathrine Charlotte; Grostøl Dietz, Andrea; Taylor, Brian R.; Drobizhev, Mikhail; Molina, Rosana S.; Aggarwal, Abhi; Rancic, Vladimir; Becker, Michael; Ballanyi, Klaus; Podgorski, Kaspar; Hirase, Hajime; Nedergaard, Maiken; Fendrych, Matyáš; Lemieux, M. Joanne; Eberl, Daniel F.; Kay, Alan R.; Campbell, Robert E.; Shen, YiPotassium ion (K+) plays a critical role as an essential electrolyte in all biological systems. Genetically-encoded fluorescent K+ biosensors are promising tools to further improve our understanding of K+-dependent processes under normal and pathological conditions. Here, we report the crystal structure of a previously reported genetically-encoded fluorescent K+ biosensor, GINKO1, in the K+-bound state. Using structure-guided optimization and directed evolution, we have engineered an improved K+ biosensor, designated GINKO2, with higher sensitivity and specificity. We have demonstrated the utility of GINKO2 for in vivo detection and imaging of K+ dynamics in multiple model organisms, including bacteria, plants, and mice.