Bioluminescence imaging captures the expression and dynamics of endogenous p21 Promoter activity in living mice and intact cells

• Autores: Tinkum, Kelsey L.; Marpegan, Luciano; White, Lynn S.; Sun, Jinwu; Herzog, Erik D.; Piwnica Worms, David; Piwnica Worms, Helen
• Año de publicación: 2011
• Idioma: inglés
• Tipo de recurso: artículo
• Materia: bioluminescence; p21; cell-cycle
• Institución: Consejo Nacional de Investigaciones Científicas y Técnicas

To interrogate endogenous p21 WAF1/CIP1 (p21) promoter activity under basal conditions and in response to various forms of stress, knock-in imaging reporter mice in which expression of firefly luciferase (FLuc) was placed under the control of the endogenous p21 promoter within the Cdkn1a gene locus were generated. Bioluminescence imaging (BLI) of p21 promoter activity was performed noninvasively and repetitively in mice and in cells derived from these mice. We demonstrated that expression of FLuc accurately reported endogenous p21 expression at baseline and under conditions of genotoxic stress and that photon flux correlated with mRNA abundance and, therefore, bioluminescence provided a direct readout of p21 promoter activity in vivo. BLI confirmed that p53 was required for activation of the p21 promoter in vivo in response to ionizing radiation. Interestingly, imaging of reporter cells demonstrated that p53 prevents the extracellular signal-regulated kinase/mitogen-activated protein kinase pathway from activating p21 expression when quiescent cells are stimulated with serum to reenter the cell cycle. In addition, low-light BLI identified p21 expression in specific regions of individual organs that had not been observed previously. This inducible p21 FLuc knock-in reporter strain will facilitate imaging studies of p53-dependent and -independent stress responses within the physiological context of the whole animal.
Fil: Tinkum, Kelsey L.. Washington University in St. Louis; Estados Unidos
Fil: Marpegan, Luciano. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Washington University in St. Louis; Estados Unidos
Fil: White, Lynn S.. Washington University in St. Louis; Estados Unidos
Fil: Sun, Jinwu. Washington University in St. Louis; Estados Unidos
Fil: Herzog, Erik D.. Washington University in St. Louis; Estados Unidos
Fil: Piwnica Worms, David. Washington University in St. Louis; Estados Unidos
Fil: Piwnica Worms, Helen. Washington University in St. Louis; Estados Unidos