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A RhoA-FRET biosensor mouse for intravital imaging in normal tissue homeostasis and disease contexts

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journal contribution
posted on 15.10.2020, 09:51 by Max Nobis, David Herrmann, Sean C Warren, Shereen Kadir, Wilfred Leung, Monica Killen, Astrid Magenau, David Stevenson, Morghan C Lucas, Nadine Reischmann, Claire Vennin, James RW Conway, Alice Boulghourjian, Anaiis Zaratzian, Andrew M Law, David Gallego-Ortega, Christopher J Ormandy, Stacey N Walters, Shane T Grey, Jacqueline Bailey, Tatyana Chtanova, Julian MW Quinn, Paul A Baldock, Peter I Croucher, Juliane P Schwarz, Agata Mrowinska, Lei Zhang, Herbert Herzog, Andrius Masedunskas, Edna C Hardeman, Peter W Gunning, Gonzalo Del Monte-Nieto, Richard P Harvey, Michael S Samuel, Marina Pajic, Ewan J McGhee, Anna-Karin E Johnsson, Owen J Sansom, Heidi CE Welch, Jennifer P Morton, Douglas Strathdee, Kurt I Anderson, Paul Timpson
The small GTPase RhoA is involved in a variety of fundamental processes in normal tissue. Spatiotemporal control of RhoA is thought to govern mechanosensing, growth, and motility of cells, while its deregulation is associated with disease development. Here, we describe the generation of a RhoA-fluorescence resonance energy transfer (FRET) biosensor mouse and its utility for monitoring real-time activity of RhoA in a variety of native tissues in vivo. We assess changes in RhoA activity during mechanosensing of osteocytes within the bone and during neutrophil migration. We also demonstrate spatiotemporal order of RhoA activity within crypt cells of the small intestine and during different stages of mammary gestation. Subsequently, we reveal co-option of RhoA activity in both invasive breast and pancreatic cancers, and we assess drug targeting in these disease settings, illustrating the potential for utilizing this mouse to study RhoA activity in vivo in real time.