The p53 protein plays a critical role in the organism resistance to the developement of cancer. The current model for p53 function suggests that it works as a specific transcription factor induced by DNA damage, that can act to induce apoptosis and/or block the cell cycle.
The aim of this research project is to develop an innovative approach to the selective manipulation of p53 function by virtue of the inducable intracellular expression of recombinant antibodies . Three anti-p53 monoclonal antibodies with known biological affects on p53 have been selected for this project.
The first part of this work will be to clone the recombinant scFV's of these antibodies. Then, a novel expression vector permitting the intranuclear expression of the recombinant antibodies (by fusing the scFV with the nucleoplasm in protein) will be constructed. The constructs will be tranfected in mouse 3T3 cells, and the function and expression of p53 will be assessed before and after induction of the synthesis of the recombinant antibodies. Functions to be assessed include the growth arrest anc the transcriptional response to DNA damage . This project would permit to study the biochemical activities of p53 required for growth arrest and apoptosis and would provide information about the affects of manipulating p53 activity in cells.