A Cancer Rainbow Mouse for the Simultaneous Assessment of Multiple Oncogenes

用于同时评估多种癌基因的癌症彩虹小鼠

基本信息

批准号：
8544451
负责人：
Marc G. Caron
金额：
$ 24.05万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-09-12 至 2015-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8544451
关键词：
Address Animal Model Biochemical Biological Biological Models Biology Breeding Cells Clinical Clone Cells Cloning Communities DNA Development Engineering Epitopes Foundations Gene Dosage Gene Transfer Techniques Generations Genes Genetic Genotype Histocompatibility Testing Image Injection of therapeutic agent Intestinal Cancer Investments Laboratories Malignant Neoplasms Malignant neoplasm of lung Methodology Methods Modeling Molecular Biology Monitor Morbidity - disease rate Mus Nature Oncogene Activation Oncogenes Outcome Patients Plasmids Population Preparation Proteins Reporter Research Research Personnel Resolution Resource Sharing Risk Site Solutions Techniques Technology Test Result Testing Tissues Transgenes Transgenic Mice Transgenic Organisms Tumor Suppressor Proteins Tumorigenicity Vertebral column anticancer research base blastocyst cancer therapy cell type cost design design and construction efficacy testing embryonic stem cell flexibility fluorescence imaging in vivo innovation innovative technologies interest malignant breast neoplasm mouse model mutant neoplastic cell new technology novel overexpression progenitor repository response skills treatment strategy tumor tumorigenesis tumorigenic user-friendly vector

项目摘要

DESCRIPTION (provided by applicant): Understanding the genetic mechanisms which underlie tumor development will provide a foundation for developing new generations of better and more effective cancer therapies. To address this fundamental issue, new technologies are needed that are superior to those currently available. Current state of the art mouse models enable testing the stochastic activation of oncogenes in a cell type specific fashion and provide a mechanism for testing targeted therapies. Despite these desirable features, their limited accessibility and feasibility precludes their implementation in most laboratories. Moreover, as the list of oncogenes continues to grow, reliable and efficient technologies are needed that can assess several oncogenes simultaneously in order to facilitate rapid analysis of tumorigenicity and their responses to therapy. This application provides a solution to all these issues by developing an innovative transgenic mouse platform (Crainbow) to rapidly, efficiently, and cost effectively create mouse models of tumorigenesis. The Crainbow technology will enable multiple user selectable oncogenes to be incorporated into a single transgenic mouse whereby each oncogene will be identifiable on the basis of a unique epitope tag and a separately expressed spectrally resolvable fluorescent protein. Using a strategy of Cre induced stochastic activation, the Crainbow technology enables the contribution of each oncogene to be assessed coincidentally in the same tissue with single cell resolution. Additionally, clonally derived tumor cell populations can be studied in any tissue and cell type by breeding to the plethora of validated cell type specific Cre mouse lines. Underlying the flexibility of this technology is a novel plasmid construct for incorporating the oncogenes of interest. The Crainbow plasmid can be easily modified for oncogene targeting using the molecular biology skills available in most all biology laboratories. Crainbow transgene mice can then be produced rapidly on site or obtained from other investigators or repositories. The utility and versatility of this Crainbow platform technology is demonstrated through the following specific aims which: (1) Establish a flexible and rapid strategy for cancer rainbow transgene design and construction, (2) Generate and validate cancer rainbow transgenic mice, & (3) Generate tissue specific tumors using cancer rainbow transgenic mice. The Cancer rainbow technology addresses in a single mouse shortcomings of current technology by providing a comprehensive method to simultaneously monitor the activity of multiple genes, and a means to also rapidly screen novel targeted cancer therapies. Consequently, the Cancer rainbow technology will provide a paradigm shift for studying tumorigenesis.

描述（由申请人提供）：了解肿瘤发展的遗传机制将为开发新一代更好、更有效的癌症疗法奠定基础。为了解决这个根本问题，需要优于现有技术的新技术。目前最先进的小鼠模型能够以细胞类型特异性方式测试癌基因的随机激活，并提供测试靶向治疗的机制。尽管有这些理想的功能，但其有限的可访问性和可行性阻碍了它们在大多数实验室中的实施。此外，随着癌基因列表的不断增加，需要可靠且高效的技术来同时评估多个癌基因，以便于快速分析致瘤性及其对治疗的反应。该应用通过开发创新的转基因小鼠平台（Crainbow）来快速、高效且经济高效地创建肿瘤发生的小鼠模型，为所有这些问题提供了解决方案。 Crainbow 技术将使多个用户可选择的癌基因被整合到单个转基因小鼠中，从而根据独特的表位标签和单独表达的光谱可分辨荧光蛋白来识别每个癌基因。 Crainbow 技术采用 Cre 诱导随机激活策略，能够在同一组织中以单细胞分辨率同时评估每个癌基因的贡献。此外，克隆来源的肿瘤通过培育大量经过验证的细胞类型特异性 Cre 小鼠系，可以研究任何组织和细胞类型的细胞群。该技术灵活性的基础是一种用于整合感兴趣的癌基因的新型质粒构建体。使用大多数生物实验室中可用的分子生物学技术，可以轻松修改 Crainbow 质粒以用于癌基因靶向。然后可以在现场快速生产或从其他研究人员或储存库获得 Crainbow 转基因小鼠。 Crainbow 平台技术的实用性和多功能性通过以下具体目标得到证明：(1) 建立灵活快速的癌症彩虹转基因设计和构建策略，(2) 生成并验证癌症彩虹转基因小鼠，以及 (3)使用癌症彩虹转基因小鼠产生组织特异性肿瘤。癌症彩虹技术通过提供一种同时监测多个基因活性的综合方法，以及一种快速筛选新型靶向癌症疗法的方法，解决了当前技术的缺点。因此，癌症彩虹技术将为研究肿瘤发生提供范式转变。