A noninvasive iRFP713 p53 reporter reveals dynamic p53 activity in response to irradiation and liver regeneration in vivo

Humpton, T. J. et al. (2022) A noninvasive iRFP713 p53 reporter reveals dynamic p53 activity in response to irradiation and liver regeneration in vivo. Science Signaling, 15(720), eabd9099. (doi: 10.1126/scisignal.abd9099) (PMID:35133863) (PMCID:PMC7612476)

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Genetically encoded probes are widely used to visualize cellular processes in vitro and in vivo. Although effective in cultured cells, fluorescent protein tags and reporters are suboptimal in vivo because of poor tissue penetration and high background signal. Luciferase reporters offer improved signal-to-noise ratios but require injections of luciferin that can lead to variable responses and that limit the number and timing of data points that can be gathered. Such issues in studying the critical transcription factor p53 have limited insight on its activity in vivo during development and tissue injury responses. Here, by linking the expression of the near-infrared fluorescent protein iRFP713 to a synthetic p53-responsive promoter, we generated a knock-in reporter mouse that enabled noninvasive, longitudinal analysis of p53 activity in vivo in response to various stimuli. In the developing embryo, this model revealed the timing and localization of p53 activation. In adult mice, the model monitored p53 activation in response to irradiation and paracetamol- or CCl4-induced liver regeneration. After irradiation, we observed potent and sustained activation of p53 in the liver, which limited the production of reactive oxygen species (ROS) and promoted DNA damage resolution. We propose that this new reporter may be used to further advance our understanding of various physiological and pathophysiological p53 responses.

Item Type:Articles
Additional Information:This work was funded by Cancer Research UK grant C596/A26855 and supported by The Francis Crick Institute, which receives its core funding from Cancer Research UK (FC001557), the United Kingdom Medical Research Council (FC001557), and the Wellcome Trust (FC001557), and the CRUK Beatson Institute, which receives its core funding from Cancer Research UK grant C596/A17196 and A31287. Additional funding for the work was provided by Cancer Research UK grant A29799 (KB). M.D.D., F.F., and L.M.C. are grateful for the support from Cancer Research UK Beatson Institute core funding (C596/A23983) and Breast Cancer Now (2019DecPR1424). T.G.B. was funded by the Wellcome Trust (grant number WT107492Z) and CRUK HUNTER Accelerator Award (grant number A26813).
Glasgow Author(s) Enlighten ID:Blyth, Professor Karen and Bryson, Miss Sheila and De Donatis, Marco and Carlin, Dr Leo and Kiourtis, Christos and Bird, Dr Thomas and Nixon, Mr Colin and Vousden, Karen and Strathdee, Mr Douglas
Authors: Humpton, T. J., Hock, A. K., Kiourtis, C., De Donatis, M., Fercoq, F., Nixon, C., Bryson, S., Strathdee, D., Carlin, L. M., Bird, T. G., Blyth, K., and Vousden, K.
College/School:College of Medical Veterinary and Life Sciences
College of Medical Veterinary and Life Sciences > School of Cancer Sciences
Journal Name:Science Signaling
Publisher:American Association for the Advancement of Science
ISSN (Online):1937-9145
Copyright Holders:Copyright © 2022 The Authors
First Published:First published in Science Signaling 15(720):eabd9099
Publisher Policy:Reproduced in accordance with the copyright policy of the publisher

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
301496HUNTER: Hepatocellular Carcinoma Expediter NetworkThomas BirdCancer Research UK (CRUK)BH172934 - C9380/A26813CS - Experimental Therapeutics