Cargando…

A non-invasive iRFP713 p53 reporter reveals dynamic p53 activity in response to irradiation and liver regeneration in vivo(†)

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 sub-optimal in vivo due to poor tissue penetration and high background signal. Luciferase reporters offer improved signal-...

Descripción completa

Detalles Bibliográficos
Autores principales: Humpton, Timothy J, Hock, Andreas K, Kiourtis, Christos, Donatis, Marco De, Fercoq, Frederic, Nixon, Colin, Bryson, Sheila, Strathdee, Douglas, Carlin, Leo M., Bird, Thomas G., Blyth, Karen, Vousden, Karen H
Formato: Online Artículo Texto
Lenguaje:English
Publicado: 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7612476/
https://www.ncbi.nlm.nih.gov/pubmed/35133863
http://dx.doi.org/10.1126/scisignal.abd9099
_version_ 1783605371500232704
author Humpton, Timothy J
Hock, Andreas K
Kiourtis, Christos
Donatis, Marco De
Fercoq, Frederic
Nixon, Colin
Bryson, Sheila
Strathdee, Douglas
Carlin, Leo M.
Bird, Thomas G.
Blyth, Karen
Vousden, Karen H
author_facet Humpton, Timothy J
Hock, Andreas K
Kiourtis, Christos
Donatis, Marco De
Fercoq, Frederic
Nixon, Colin
Bryson, Sheila
Strathdee, Douglas
Carlin, Leo M.
Bird, Thomas G.
Blyth, Karen
Vousden, Karen H
author_sort Humpton, Timothy J
collection PubMed
description 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 sub-optimal in vivo due to 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 non-invasive, 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 CCl(4)- 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.
format Online
Article
Text
id pubmed-7612476
institution National Center for Biotechnology Information
language English
publishDate 2022
record_format MEDLINE/PubMed
spelling pubmed-76124762022-03-07 A non-invasive iRFP713 p53 reporter reveals dynamic p53 activity in response to irradiation and liver regeneration in vivo(†) Humpton, Timothy J Hock, Andreas K Kiourtis, Christos Donatis, Marco De Fercoq, Frederic Nixon, Colin Bryson, Sheila Strathdee, Douglas Carlin, Leo M. Bird, Thomas G. Blyth, Karen Vousden, Karen H Sci Signal Article 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 sub-optimal in vivo due to 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 non-invasive, 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 CCl(4)- 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. 2022-02-08 2022-02-08 /pmc/articles/PMC7612476/ /pubmed/35133863 http://dx.doi.org/10.1126/scisignal.abd9099 Text en https://creativecommons.org/licenses/by/4.0/This work is licensed under a CC BY 4.0 (https://creativecommons.org/licenses/by/4.0/) International license.
spellingShingle Article
Humpton, Timothy J
Hock, Andreas K
Kiourtis, Christos
Donatis, Marco De
Fercoq, Frederic
Nixon, Colin
Bryson, Sheila
Strathdee, Douglas
Carlin, Leo M.
Bird, Thomas G.
Blyth, Karen
Vousden, Karen H
A non-invasive iRFP713 p53 reporter reveals dynamic p53 activity in response to irradiation and liver regeneration in vivo(†)
title A non-invasive iRFP713 p53 reporter reveals dynamic p53 activity in response to irradiation and liver regeneration in vivo(†)
title_full A non-invasive iRFP713 p53 reporter reveals dynamic p53 activity in response to irradiation and liver regeneration in vivo(†)
title_fullStr A non-invasive iRFP713 p53 reporter reveals dynamic p53 activity in response to irradiation and liver regeneration in vivo(†)
title_full_unstemmed A non-invasive iRFP713 p53 reporter reveals dynamic p53 activity in response to irradiation and liver regeneration in vivo(†)
title_short A non-invasive iRFP713 p53 reporter reveals dynamic p53 activity in response to irradiation and liver regeneration in vivo(†)
title_sort non-invasive irfp713 p53 reporter reveals dynamic p53 activity in response to irradiation and liver regeneration in vivo(†)
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7612476/
https://www.ncbi.nlm.nih.gov/pubmed/35133863
http://dx.doi.org/10.1126/scisignal.abd9099
work_keys_str_mv AT humptontimothyj anoninvasiveirfp713p53reporterrevealsdynamicp53activityinresponsetoirradiationandliverregenerationinvivo
AT hockandreask anoninvasiveirfp713p53reporterrevealsdynamicp53activityinresponsetoirradiationandliverregenerationinvivo
AT kiourtischristos anoninvasiveirfp713p53reporterrevealsdynamicp53activityinresponsetoirradiationandliverregenerationinvivo
AT donatismarcode anoninvasiveirfp713p53reporterrevealsdynamicp53activityinresponsetoirradiationandliverregenerationinvivo
AT fercoqfrederic anoninvasiveirfp713p53reporterrevealsdynamicp53activityinresponsetoirradiationandliverregenerationinvivo
AT nixoncolin anoninvasiveirfp713p53reporterrevealsdynamicp53activityinresponsetoirradiationandliverregenerationinvivo
AT brysonsheila anoninvasiveirfp713p53reporterrevealsdynamicp53activityinresponsetoirradiationandliverregenerationinvivo
AT strathdeedouglas anoninvasiveirfp713p53reporterrevealsdynamicp53activityinresponsetoirradiationandliverregenerationinvivo
AT carlinleom anoninvasiveirfp713p53reporterrevealsdynamicp53activityinresponsetoirradiationandliverregenerationinvivo
AT birdthomasg anoninvasiveirfp713p53reporterrevealsdynamicp53activityinresponsetoirradiationandliverregenerationinvivo
AT blythkaren anoninvasiveirfp713p53reporterrevealsdynamicp53activityinresponsetoirradiationandliverregenerationinvivo
AT vousdenkarenh anoninvasiveirfp713p53reporterrevealsdynamicp53activityinresponsetoirradiationandliverregenerationinvivo
AT humptontimothyj noninvasiveirfp713p53reporterrevealsdynamicp53activityinresponsetoirradiationandliverregenerationinvivo
AT hockandreask noninvasiveirfp713p53reporterrevealsdynamicp53activityinresponsetoirradiationandliverregenerationinvivo
AT kiourtischristos noninvasiveirfp713p53reporterrevealsdynamicp53activityinresponsetoirradiationandliverregenerationinvivo
AT donatismarcode noninvasiveirfp713p53reporterrevealsdynamicp53activityinresponsetoirradiationandliverregenerationinvivo
AT fercoqfrederic noninvasiveirfp713p53reporterrevealsdynamicp53activityinresponsetoirradiationandliverregenerationinvivo
AT nixoncolin noninvasiveirfp713p53reporterrevealsdynamicp53activityinresponsetoirradiationandliverregenerationinvivo
AT brysonsheila noninvasiveirfp713p53reporterrevealsdynamicp53activityinresponsetoirradiationandliverregenerationinvivo
AT strathdeedouglas noninvasiveirfp713p53reporterrevealsdynamicp53activityinresponsetoirradiationandliverregenerationinvivo
AT carlinleom noninvasiveirfp713p53reporterrevealsdynamicp53activityinresponsetoirradiationandliverregenerationinvivo
AT birdthomasg noninvasiveirfp713p53reporterrevealsdynamicp53activityinresponsetoirradiationandliverregenerationinvivo
AT blythkaren noninvasiveirfp713p53reporterrevealsdynamicp53activityinresponsetoirradiationandliverregenerationinvivo
AT vousdenkarenh noninvasiveirfp713p53reporterrevealsdynamicp53activityinresponsetoirradiationandliverregenerationinvivo