Core A: Molecular Imaging Reporter Core (MIRC)

核心 A：分子成像报告核心 (MIRC)

基本信息

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
项目类别：
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

项目摘要

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.分子成像报告基因核心（MIRC）分子成像记者核心是一种提供专业知识，材料和的中央设施在设计和执行分子成像的生物学方面的协作协助。如果一个人确定代表华盛顿大学ICMIC创新本质的核心，它将成为分子成像报告基因核心。 MIRC为具有广泛范围的调查人员服务分子生物学，细胞培养和动物实验方面的资源和经验。最大的该核心的重要活动是发现和传播我们的新型分子成像试剂和遗传编码的记者向我们机构中的调查人员，其他P50计划网站以及世界各地的癌症生物学和成像研究人员。该核心的发现研究提供研究社区：1）基于蛋白质蛋白的新型宠物和生物发光记者基于修饰的两个杂交转录策略的体内相互作用，2）新型萤火虫荧光素酶蛋白质碎片互补策略用于体内蛋白质 - 蛋白质相互作用的实时成像，3）新颖的融合蛋白策略在体内成像蛋白酶体功能，4）创新平台策略实时询问泛素诱导的配体调节蛋白的降解（例如，KB，P-catenin和cdc25a），5）第二代融合记者和三模式记者多模式成像（PET/生物发光/荧光），6）设计方便的矢量来克隆并表达点击甲虫荧光素酶，萤火虫荧光素酶，Renilla荧光素酶，MTHSV1-TK，MTSSTR-2和其他用于各种成像应用的，以及7）产生转基因和敲击分子成像记者小鼠（例如GAL4-FLUC，P21-FLUC，ROSA26-LSL-CGR-MGFP）。确实，这个核心一直是我们最有生产力的人之一，全面的活动，发现和发展新的记者并影响广泛的范围全世界的研究计划。华盛顿大学的许多调查员社区和外部机构直接收到了Wu Mirc的材料和支持在进度报告涵盖的5年期间。这些活动包括新计划持续协作项目以及试点项目，这些项目现在将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。总体而言，截至2006年6月，我们已经分发了分子成像报告基因和细胞的收集到数十个WU研究者以及86 全世界的调查人员。其他几个P50 ICMIC机构也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的研究人员（裂开萤火虫荧光素酶， Ub-Fluc质粒，IKB-FLUC和对照质粒）；斯坦福大学ICMIC的调查员（IKB-FLUC，， FLUC载体，表达IKB-FLUC的稳定记者细胞，鞘翅类类似物）；调查人员密歇根大学ICMIC（分裂萤火虫荧光素酶，UB-Fluc质粒）；和哈佛的调查员萤火虫荧光素酶，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-Firefly荧光素酶融合构建体，多泛素化 - Firefly Luciferase，突变NLS-SR39HSV1-TK-EGFP融合记者和GAL4-FIREFLY荧光素酶记者。知名期刊的出版物（例如PNAS 2006，103：1313-1318）已经出现在文献以我们作为各自项目的这些分子成像试剂来源。