Development of a p53 responsive GFP reporter; identification of live cellswith p53 activity

p53 is among the most intensely studied human proteins because of its vital role as the prototype tumor suppressor. As a result, there are widespread applications for p53 functional analysis in biotechnology as it relates to cancer research. p53 is a potent sequence specific transcription factor, wh ich induces the expression of a number of genes whose products mediate cell cycle arrest and apoptosis. Because the tumor suppressor activity of p53 i s dependent on its transcription regulatory function, we have undertaken to develop a p53-responsive green fluorescent protein reporter strategy to en able the identification of live cells containing p53 transcriptional transa ctivation activity. We demonstrate within the use of GFP fluorescence to mo nitor both endogenous and plasmid derived p53 biochemical and biological ac tivity. Identifying live cells with p53 activity through GFP fluorescence w ill have wide application for both in vitro and in vivo studies of the p53 tumor suppressor protein. (C) 2000 Elsevier Science B.V. All rights reserve d.