Core A: Molecular Imaging Reporter Core (MIRC)

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A.3.I. Molecular Imaging Reporter Core (MIRC) The Molecular Imaging Reporter Core is a central facility providing expertise, materials and collaborative assistance for design and execution of biological aspects of molecular imaging. If one was to identify the one Core that represented the essence of ICMIC innovation at Washington University, it would be the Molecular Imaging Reporter Core. The MIRC serves investigators possessing a wide range of resources and experience in molecular biology, cell culture, and animal experimentation. One of the most important activities of this core is discovery research and dissemination of our novel molecular imaging reagents and genetically-encoded reporters to investigators within our institution, to other P50 program sites and to cancer biology and imaging investigators throughout the world. Discovery research in this Core provided the research community with: 1) Novel PET- and bioluminescence-based reporters of proteinprotein interactions in vivo based on modified two-hybrid transcriptional strategies, 2) Novel firefly luciferase protein fragment complementation strategies for real-time imaging of protein-protein interactions in vivo, 3) Novel fusion protein strategies for imaging proteasome function in vivo, 4) Innovative platform strategies to interrogate ubiquitin-induced degradation of ligand-regulated proteins in signaling pathways in real time (e.g., kB, p-catenin and Cdc25A), 5) Second generation fusion reporters and triple-modality reporters for multi-modality imaging (PET/bioluminescence/ fluorescence), 6) Engineered convenient vectors for cloning and expression of click beetle luciferases, firefly luciferase, Renilla luciferase, mtHSV1-TK, mtSSTR-2 and others for a variety of imaging applications, and 7) Production of transgenic and knock-in molecular imaging reporter mice (e.g., Gal4-Fluc, p21-Fluc, ROSA26-LSL-CGR-mGFP). Indeed, this Core has been and continues to be one of our most productive and comprehensive activities, discovering and developing novel reporters and impacting a broad range of research programs throughout the world. Many investigators within the Washington University community as well as outside institutions have directly received material and support from the WU MIRC during the 5 year period covered by the progress report. These activities include new initiatives, continuation collaborative projects as well as pilot projects that now extend our reach far beyond the focus of the original projects proposed for the Center Program. Overall, as of June 2006, we have distributed our collection of molecular imaging reporter reagents and cells to dozens of WU investigators as well as 86 investigators throughout the world. Several other P50 ICMIC institutions have requested and received our cells and reagents, including investigators at the Johns Hopkins University ICMIC (split firefly luciferase, Ub-FLuc plasmid, IkB-FLuc and control plasmids); investigators at the Stanford University ICMIC (IkB-FLuc, FLuc vectors, stable reporter cells expressing IkB-FLuc, coelenterazine analogues); investigators at the University of Michigan ICMIC (split firefly luciferase, Ub-FLuc plasmid); and investigators at Harvard (split firefly luciferase, IkB-FLuc). Our most popular reagents (implying high impact) include plasmids encoding our luciferase complementation fragments (split luciferase), IkB-firefly luciferase fusion construct, polyubiquitinated- firefly luciferase, mutant NLS-sr39HSV1-TK-EGFP fusion reporter, and Gal4-firefly luciferase reporter. Publications in high profile journals (e.g., PNAS 2006, 103:1313-1318) have already appeared in the literature citing us as the source of these molecular imaging reagents for their respective projects.