한빛사 논문
Jihoon Kim1, Sangkyu Lee2, Kanghoon Jung3, Won Chan Oh3,4, Nury Kim2, Seungkyu Son1, YoungJu Jo5, Hyung-Bae Kwon3,6 & Won Do Heo1,2,7
1 Department of Biological Sciences, Korea Advanced Institute of Science and Technology (KAIST), Daejeon 34141, Republic of Korea. 2 Center for Cognition and Sociality, Institute for Basic Science (IBS), Daejeon 34126, Republic of Korea. 3 Max Planck Florida Institute for Neuroscience (MPFI), Jupiter, FL 33458, USA. 4 Department of Pharmacology, University of Colorado School of Medicine, Anschutz Medical Campus, Aurora, CO 80045, USA. 5 Department of Physics, Korea Advanced Institute of Science and Technology (KAIST), Daejeon 34141, Republic of Korea. 6 Max Planck Institute of Neurobiology, Martinsried 82152, Germany. 7 KAIST Institute for the BioCentury, Korea Advanced Institute of Science and Technology (KAIST), Daejeon 34141, Republic of Korea. These authors contributed equally: Jihoon Kim, Sangkyu Lee, Kanghoon Jung.
Correspondence to Hyung-Bae Kwon or Won Do Heo
Abstract
Ras and Rho small GTPases are critical for numerous cellular processes including cell division, migration, and intercellular communication. Despite extensive efforts to visualize the spatiotemporal activity of these proteins, achieving the sensitivity and dynamic range necessary for in vivo application has been challenging. Here, we present highly sensitive intensiometric small GTPase biosensors visualizing the activity of multiple small GTPases in single cells in vivo. Red-shifted sensors combined with blue light-controllable optogenetic modules achieved simultaneous monitoring and manipulation of protein activities in a highly spatiotemporal manner. Our biosensors revealed spatial dynamics of Cdc42 and Ras activities upon structural plasticity of single dendritic spines, as well as a broad range of subcellular Ras activities in the brains of freely behaving mice. Thus, these intensiometric small GTPase sensors enable the spatiotemporal dissection of complex protein signaling networks in live animals.
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