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

• 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
• 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

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）：了解肿瘤发展构成的遗传机制将为发展新一代更好，更有效的癌症疗法提供基础。为了解决这个基本问题，需要新技术优于当前可用的技术。当前的ART小鼠模型可以以细胞类型的方式测试癌基因的随机激活，并提供了测试靶向疗法的机制。尽管有这些理想的功能，但它们的可访问性和可行性有限，排除了它们在大多数实验室中的实施。此外，随着肿瘤基因的列表不断增长，需要可靠，有效的技术，可以同时评估几种发病基因，以促进对肿瘤性的快速分析及其对治疗的反应。该应用程序通过开发创新的转基因小鼠平台（CRAINBOW）来快速，有效地创建肿瘤发生的小鼠模型，从而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的致癌基因掺入单个转基因小鼠中，从而根据唯一的表位标签和单独表达的频谱可分离荧光蛋白来识别每种癌基因。使用CRE诱导的随机激活策略，Crainbow技术使每种癌基因的贡献都可以在同一组织中以单细胞分辨率进行偶然评估。另外，克隆衍生的肿瘤 可以通过在任何组织和细胞类型中研究细胞群来通过繁殖过多的经过验证的细胞类型特异性CRE小鼠系。这项技术的灵活性是一种新型的质粒构造，用于融合感兴趣的肿瘤基因。使用大多数所有生物学实验室中可用的分子生物学技能，可以轻松地修改Crainbow质粒，以靶向癌基因。然后可以在现场快速生产Crainbow转基因小鼠，也可以从其他研究人员或存储库中获得。通过以下特定目的证明了这种Crainbow平台技术的实用性和多功能性，（1）为癌症彩虹转基因设计和构建建立灵活而快速的策略，（2）生成和验证癌症彩虹转基因小鼠，＆（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全面的方法来同时监测多个基因的活性以及一种迅速筛查新颖的靶向癌症疗法的方法来解决当前技术的单一小鼠缺点。因此，癌症彩虹技术将为研究肿瘤发生提供范式转移。