Core A: Molecular Imaging Reporter Core (MIRC)

A.3.I.分子成像报告核心(MIRC) 分子成像报告核心是一个中央设施，提供专业知识、材料和 协作性协助设计和执行分子成像的生物学方面。如果有人要 找出代表华盛顿大学ICMIC创新本质的一个核心，它将 成为分子成像报道核心。MIRC为拥有广泛 在分子生物学、细胞培养和动物实验方面的资源和经验。其中最 这个核心的重要活动是发现、研究和传播我们的新型分子成像 试剂和基因编码记者到我们机构的研究人员，到其他P50计划站点 以及世界各地的癌症生物学和成像研究人员。在这个核心中的发现研究 为研究界提供了：1)基于PET和生物发光的新型蛋白质报告 基于改进双杂交转录策略的体内相互作用，2)新的萤火虫荧光素酶 蛋白质片段互补策略用于活体蛋白质相互作用的实时成像，3) 用于体内蛋白酶体功能成像的新的融合蛋白策略，4)创新的平台策略 实时检测泛素诱导的信号通路中配体调节蛋白的降解 (例如，kB、p-catenin和CDc25A)，5)第二代融合记者和用于 多模式成像(PET/生物发光/荧光)，6)构建了方便克隆的载体 以及甲虫荧光素酶、萤火虫荧光素酶、雷尼拉荧光素酶、mtHSV1-TK、mtSSTR-2和 其他用于各种成像应用，以及7)转基因和嵌合分子成像的生产 报告小鼠(例如Gal4-Fluc、p21-Fluc、rosa26-LSL-CGR-mGFP)。 事实上，这一核心一直是并将继续是我们最具生产力和 活动全面，发现和发展新颖的记者，影响广泛 世界各地的研究项目。华盛顿大学内部的许多调查人员 社区以及外部机构直接获得了来自吴MIRC的材料和支持 在进度报告所述的5年期间。这些活动包括新的倡议， 持续的合作项目以及试点项目，这些项目现在远远超出了我们的关注范围 为中心计划提出的原始项目。总体而言，截至2006年6月，我们已经分发了我们的 收集分子成像报告试剂和细胞给数十名吴调查人员以及86名 世界各地的调查人员。其他几个P50 ICMIC机构已经要求并收到了我们的 细胞和试剂，包括约翰霍普金斯大学ICMIC(分裂萤火虫荧光素酶， UB-flc质粒、IKB-flc和对照质粒)；斯坦福大学ICMIC的研究人员(IKB-flc， Flc载体，表达IKB-flc的稳定报告细胞，coelenterazine类似物)；研究人员在 密歇根大学ICMIC(分裂萤火虫荧光素酶，Ub-flc质粒)；哈佛大学研究人员(分裂 萤火虫荧光素酶(IKB-flc)。我们最受欢迎的试剂(意味着高影响力)包括编码的质粒 我们的荧光素酶互补片段(分裂的荧光素酶)，IKB-萤火虫荧光素酶融合构建，多泛素化- 萤火虫荧光素酶、突变体NLS-sr39HSV1-TK-EGFP融合报告基因和Gal4-萤火虫荧光素酶 记者。知名期刊(例如，PNAS 2006,103：1313-1318)中的出版物已经出现在 这些文献引用了我们作为这些分子成像试剂的来源，用于他们各自的项目。

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.