| Autor: |
Humpton TJ; The Francis Crick Institute, London NW1 1AT, UK.; Cancer Research UK Beatson Institute, Glasgow G61 1BD, UK., Hock AK; Cancer Research UK Beatson Institute, Glasgow G61 1BD, UK., Kiourtis C; Cancer Research UK Beatson Institute, Glasgow G61 1BD, UK.; Institute of Cancer Sciences, University of Glasgow, Glasgow G61 1QH, UK., De Donatis M; Cancer Research UK Beatson Institute, Glasgow G61 1BD, UK.; Institute of Cancer Sciences, University of Glasgow, Glasgow G61 1QH, UK., Fercoq F; Cancer Research UK Beatson Institute, Glasgow G61 1BD, UK., Nixon C; Cancer Research UK Beatson Institute, Glasgow G61 1BD, UK., Bryson S; Cancer Research UK Beatson Institute, Glasgow G61 1BD, UK., Strathdee D; Cancer Research UK Beatson Institute, Glasgow G61 1BD, UK., Carlin LM; Cancer Research UK Beatson Institute, Glasgow G61 1BD, UK.; Institute of Cancer Sciences, University of Glasgow, Glasgow G61 1QH, UK., Bird TG; Cancer Research UK Beatson Institute, Glasgow G61 1BD, UK.; MRC Centre for Inflammation Research, The Queen's Medical Research Institute, University of Edinburgh, Edinburgh EH16 4TJ, UK., Blyth K; Cancer Research UK Beatson Institute, Glasgow G61 1BD, UK.; Institute of Cancer Sciences, University of Glasgow, Glasgow G61 1QH, UK., Vousden KH; The Francis Crick Institute, London NW1 1AT, UK. |
| Abstrakt: |
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 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. |
