A Cancer Rainbow Mouse for the Simultaneous Assessment of Multiple Oncogenes

描述(申请人提供)：了解肿瘤发展背后的遗传机制将为开发更好和更有效的新一代癌症治疗方法提供基础。为了解决这一根本问题，需要比现有技术更优越的新技术。目前最先进的小鼠模型能够以细胞类型特定的方式测试癌基因的随机激活，并提供用于测试靶向治疗的机制。尽管有这些令人向往的特点，但它们有限的可及性和可行性阻碍了它们在大多数实验室的实施。此外，随着致癌基因名单的持续增长，需要可靠和有效的技术来同时评估几个致癌基因，以便于快速分析肿瘤的致瘤性及其对治疗的反应。该应用程序通过开发一种创新的转基因小鼠平台(Cainrow)来快速、高效、经济高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。彩虹技术将使多个用户可选择的癌基因被整合到一个转基因小鼠中，从而每个癌基因将根据一个独特的表位标签和一个单独表达的光谱可分辨荧光蛋白来识别。利用CRE诱导随机激活的策略，彩虹技术使每个癌基因的贡献能够在同一组织中以单细胞分辨率得到一致评估。此外，克隆性肿瘤 在任何组织和细胞类型中，通过繁殖过多的经过验证的细胞类型特定的Cre小鼠品系，可以研究细胞群体。这项技术的灵活性背后是一种新的质粒构建，用于整合感兴趣的癌基因。利用大多数生物实验室提供的分子生物学技术，可以很容易地为癌基因打靶而修改Cainrow质粒。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彩虹转基因小鼠。通过以下具体目标论证了该彩虹平台技术的实用性和通用性：(1)建立灵活、快速的癌症彩虹转基因设计和构建策略，(2)产生和验证癌症彩虹转基因小鼠，(3)利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动，并提供了一种快速筛选新的靶向癌症疗法的手段，从而在单个鼠标中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一个范式转换。

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.