In Vivo Imaging of MDR1A Gene Expression

摘要

Our experience studying the MDRl gene prompted us to initiate work on a novel animal model to study MDRl/mdrl gene expression under a variety of normal and breast cancer-related physiological conditions. With the advent of new bioimaging technology and the advancement of efficient gene targeting strategies, we found an opportunity to apply these state-of-the-art molecular tools to our problem. The work performed with the support of this grant has enabled us to; (1) engineere a targeting vector to allow insertion of a reporter (luciferase or HSV-tk) into the genomic locus of the mouse mdrla gene; (2) create mouse embryonic stem cells in which a gene replacement/knock-in strategy was used to insert luciferase into the mouse mdrl a genomic locus; (3) demonstrate that luciferase expression in these cells requires Cre recombinase to bring luciferase in-frame with the translational start site of the mdrla gene product; (4) show that the recombined configuration of mdrl/LUC, in its cDNA form, encodes a functional protein with luciferase activity, and (5) create both founder and Cre-recombinase expressing mouse strains for use in vivo imaging experiments. Work performed to date has proved the feasibility of this approach. However, further refinements to the model are required.