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提供专业知识、材料和 分子成像生物学方面的设计和执行的协作协助。如果有人要 确定代表华盛顿大学 ICMIC 创新本质的一个核心，它将 成为分子成像报告核心。 MIRC 为拥有广泛范围的调查人员提供服务 分子生物学、细胞培养和动物实验方面的资源和经验。最有之一 该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给我们机构内的研究人员以及其他 P50 计划站点 以及世界各地的癌症生物学和影像研究人员。该核心的发现研究 为研究界提供： 1) 新型基于 PET 和生物发光的蛋白质报告基因 基于改良的双杂交转录策略的体内相互作用，2）新型萤火虫荧光素酶 用于体内蛋白质-蛋白质相互作用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）创新平台策略 实时探究信号通路中泛素诱导的配体调节蛋白的降解 （例如，kB、p-catenin 和 Cdc25A），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) 设计方便的克隆载体 以及点击甲虫荧光素酶、萤火虫荧光素酶、海肾荧光素酶、mtHSV1-TK、mtSSTR-2 和的表达 其他用于各种成像应用，以及 7) 转基因和基因敲入分子成像的生产 报告小鼠（例如 Gal4-Fluc、p21-Fluc、ROSA26-LSL-CGR-mGFP）。 事实上，这个核心一直并将继续是我们最具生产力和效率的核心之一。 综合活动，发现和培养新颖的记者并影响广泛的范围 世界各地的研究项目。华盛顿大学内的许多研究人员 社区和外部机构直接获得了 WU MIRC 的物质和支持 进度报告涵盖的 5 年期间。这些活动包括新举措、 持续的合作项目以及试点项目现在将我们的影响范围远远超出了焦点 为中心计划提议的原始项目。总体而言，截至 2006 年 6 月，我们已经分发了 为数十名 WU 研究人员以及 86 名研究人员收集分子成像报告试剂和细胞 世界各地的调查人员。其他几家 P50 ICMIC 机构已请求并收到了我们的 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-FLuc 质粒、IkB-FLuc 和对照质粒）；斯坦福大学 ICMIC (IkB-FLuc, FLuc载体、表达IkB-FLuc的稳定报告细胞、腔肠素类似物）；调查人员在 密歇根大学ICMIC（裂解萤火虫荧光素酶，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）包括编码质粒 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、多泛素化- 萤火虫荧光素酶、突变型 NLS-sr39HSV1-TK-EGFP 融合报告基因和 Gal4-萤火虫荧光素酶 记者。知名期刊上的出版物（例如，PNAS 2006, 103:1313-1318）已出现在 文献引用我们作为其各自项目的这些分子成像试剂的来源。