Core A: Molecular Imaging Reporter Core (MIRC)

核心 A：分子成像报告核心 (MIRC)

• 批准号: 7287034
• 负责人: David Piwnica-Worms
• 金额: $ 27.55万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-28 至 2011-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7287034
• 关键词: Animal Experimentation; Animals; Archives; Biological; Bioluminescence; Breeding; Cancer Biology; Cataloging; Catalogs; Cell Line; Cells; Cellular biology; Chimeric Proteins; Cloning; Cloning Vectors; Collection; Communities; Computer software; Core Facility; Culture Media; Cultured Cells; Databases; Date of birth; Doctor of Philosophy; Engineered Gene; Engineering; Environment; Equipment and supply inventories; Experimental Designs; Firefly Luciferases; Fluorescence; Freezing; Gene Expression; Gene Expression Regulation; Generations; Genetic Transduction; Image; Institution; Journals; Knock-in Mouse; Label; Left; Ligands; Link; Literature; Luciferases; Mammalian Cell; Maps; Michigan; Modality; Molecular; Molecular Biology; Mus; Numbers; Online Systems; Order Coleoptera; Parents; Pilot Projects; Plasmids; Positioning Attribute; Positron-Emission Tomography; Production; Progress Reports; Protein Fragment; Proteins; Protocols documentation; Publications; Published Comment; Radio; Range; Reagent; Records; Renilla Luciferases; Reporter; Reporter Genes; Reporting; Research; Research Activity; Research Personnel; Research Project Grants; Resources; Scientist; Services; Signal Pathway; Site; Source; Specialized Center; Standards of Weights and Measures; System; TNFRSF5 gene; Time; Training; Transgenes; Transgenic Animals; Transgenic Organisms; Ubiquitin; Universities; Vertebral column; Vial device; Virus; Washington; Xenograft procedure; analog; animal care; base; coelenterazine; design; dissemination research; established cell line; experience; expression cloning; in vitro Assay; in vivo; in vivo Cellular and Molecular Imaging Centers; innovation; interest; molecular imaging; multicatalytic endopeptidase complex; mutant; novel; programs; promoter; protein protein interaction; repository; research study; sex; vector; yeast two hybrid system

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.

A.3.I.分子成像报告核心（MIRC） 分子成像报告核心是一个中心设施，提供专业知识，材料和 为分子成像的生物学方面的设计和执行提供协作协助。如果要 确定一个代表华盛顿大学ICMIC创新精髓的核心， 成为分子成像报告核心。MIRC为调查人员提供广泛的 在分子生物学、细胞培养和动物实验方面的资源和经验。一个最 该中心的重要活动是发现、研究和传播我们的新型分子成像技术 试剂和基因编码的报告基因提供给我们机构内的研究者，以及其他P50项目研究中心 以及全世界的癌症生物学和成像研究人员。本核心的发现研究 为研究界提供了：1）基于PET和生物发光的蛋白质蛋白质报告分子 2）新型萤火虫荧光素酶 用于体内蛋白质-蛋白质相互作用的实时成像的蛋白质片段互补策略，3） 用于体内蛋白酶体功能成像的新型融合蛋白策略，4）用于体内蛋白酶体功能成像的创新平台策略， 真实的实时地询问泛素诱导的信号通路中配体调节蛋白的降解 (e.g., kB、β-连环蛋白和Cdc 25 A），5）第二代融合报告基因和三模态报告基因， 多模态成像（PET/生物发光/荧光），6）用于克隆的工程化方便载体 以及表达卡克甲虫荧光素酶、萤火虫荧光素酶、海肾荧光素酶、mtHSV 1-TK、mtSSTR-2和 其它用于各种成像应用，和7）产生转基因和敲入分子成像 报告小鼠（例如，Gal4-Fluc，p21-Fluc，ROSA 26-LSL-CGR-mGFP）。 事实上，这一核心一直是并将继续是我们最富有成效和 综合活动，发现和发展新的记者，影响广泛 在世界各地的研究项目。华盛顿大学的许多调查人员 社区以及外部机构直接从WU MIRC获得材料和支持 在进度报告所涵盖的五年期间。这些活动包括新的倡议， 继续合作项目以及试点项目，现在我们的影响力远远超出了重点 为中心计划提出的原始项目。总的来说，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，以几十个吴研究人员以及86 全世界的调查员。其他几个P50 ICMIC机构已请求并收到我们的 细胞和试剂，包括约翰霍普金斯大学ICMIC的研究者（分裂萤火虫荧光素酶， Ub-FLuc质粒，IkB-FLuc和对照质粒）;斯坦福大学ICMIC的研究人员（IkB-FLuc， FLuc载体，表达IkB-FLuc的稳定报告细胞，腔肠素类似物）; 密歇根大学ICMIC（分裂萤火虫荧光素酶，Ub-FLuc质粒）和哈佛大学的研究人员（分裂 萤火虫荧光素酶，IkB-FLuc）。我们最受欢迎的试剂（意味着高影响力）包括质粒编码 我们的荧光素酶互补片段（分裂荧光素酶），IkB-萤火虫荧光素酶融合构建体，多聚泛素化- 萤火虫荧光素酶、突变型NLS-sr 39 HSV 1-TK-EGFP融合报告基因和Gal 4-萤火虫荧光素酶 记者.在高知名度期刊上发表的文章（例如，PNAS 2006，103：1313-1318）已经出现在 文献引用我们作为这些分子成像试剂的来源，用于各自的项目。