Intensiometric biosensors visualize the activity of multiple small GTPases in vivo

Jihoon Kim, Sangkyu Lee, Kanghoon Jung, Won Chan Oh, Nury Kim, Seungkyu Son, YoungJu Jo, Hyung-Bae Kwon () and Won Do Heo ()
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Jihoon Kim: Korea Advanced Institute of Science and Technology (KAIST)
Sangkyu Lee: Center for Cognition and Sociality, Institute for Basic Science (IBS)
Kanghoon Jung: Max Planck Florida Institute for Neuroscience (MPFI)
Won Chan Oh: Max Planck Florida Institute for Neuroscience (MPFI)
Nury Kim: Center for Cognition and Sociality, Institute for Basic Science (IBS)
Seungkyu Son: Korea Advanced Institute of Science and Technology (KAIST)
YoungJu Jo: Korea Advanced Institute of Science and Technology (KAIST)
Hyung-Bae Kwon: Max Planck Florida Institute for Neuroscience (MPFI)
Won Do Heo: Korea Advanced Institute of Science and Technology (KAIST)

Nature Communications, 2019, vol. 10, issue 1, 1-11

Abstract: 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.

Date: 2019
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DOI: 10.1038/s41467-018-08217-3

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