Instituting innovative CRISPR/Cas9 technology at an NCI Comprehensive Cancer Center transgenic facility

项目摘要/摘要 基因工程动物模型对于了解基因如何影响癌症至关重要 易感性、发展和进展。因此，动物模型核心服务对于 综合癌症中心(CCC)。MD Anderson的基因工程小鼠设施(GEMF) 癌症中心由Guillermina Lozano博士于1988年创立，目的是在一个 努力评估被确定为可能的肿瘤抑制和激活基因。与许多基本的和 MD Anderson的翻译研究项目是基于对癌症的基因分析，我们的研究 教职员工必须能够使用强大的转基因设施。GEMF提供了一个通用的状态源 为洛扎诺博士和其他MD Anderson研究人员提供的尖端专业知识和技术技能 他们的转基因小鼠模型。由于每个鼠标项目的个别性质，GEMF 为所有机构研究人员提供全面的动物模型服务，而不考虑先前的专业知识。 GEMF已经为150多个不同的癌症中心成员提供了服务。小鼠模型 由GEMF产生的物质已被160多种出版物报道的研究中使用。简·帕克博士- Thornburg(本文作者是MD Anderson的GEMF联席主任)提供基本的个性化服务 包括：1)为研究人员的特定需求提供最佳动物模型的咨询；2)生成 商定的模型，以及3)如何照顾和评估所产生的模型的培训。我把它带进来测试 供MD Anderson调查人员使用的尖端技术。预计我将制定新的战略和 满足我们教职员工需求的技术。令人兴奋的新CRISPR/CAS9技术目前正在 在GEMF中使用、评估和改进。为现场调查人员执行的工作(Richard Behringer，Gigi Lozano)对这项技术提出了许多见解，包括描述了 产生F0动物，并建立新的方法来提高精确靶向率(同源性驱动 修复)。已经建立了合作来开发和测试新的方法来生成条件 突变。新的具有不同序列特异性的CaS酶和那些提供更忠实活性的酶将 接受测试。我们的结果将在验证后与其他转基因机构广泛共享。我们高度重视 参与开发和标准化MD Anderson和 世界各地的转基因设施。

Project Summary/Abstract Genetically engineered animal models are essential for understanding how genes can affect cancer susceptibility, development, and progression. For that reason, an animal model core service is essential for a Comprehensive Cancer Center (CCC). The Genetically Engineered Mouse Facility (GEMF) at MD Anderson Cancer Center was begun in 1988 by Dr. Guillermina Lozano to generate genetically modified animals in an effort to evaluate genes identified as possible tumor suppressors and activators. As many basic and translational research programs at MD Anderson are based on a genetic analyses of cancer, our research faculty must be able to access a strong transgenic facility. The GEMF provides a common source of state-of- the-art expertise and technical skills for Dr. Lozano and other MD Anderson investigators for the generation of their genetically engineered mouse models. Because of the individual nature of each mouse project, the GEMF provides comprehensive animal model services to all institutional investigators, regardless of prior expertise. The GEMF has provided services to more than 150 different Cancer Center members. Mouse models generated by the GEMF have been used in research reported in more than 160 publications. Dr. Jan Parker- Thornburg (this author, the co-Director of the GEMF at MD Anderson) provides essential individualized service including: 1) consultation on optimal animal models for an investigator's specific needs, 2) generation of the agreed-upon model, and 3) training in how to care for and evaluate the models produced. I bring in and test cutting-edge technologies for use by MD Anderson investigators. I am expected to develop new strategies and technologies to meet the needs of our faculty. Exciting new CRISPR/Cas9 technologies are currently being used, assessed, and refined in the GEMF. Work performed for on-site investigators (Richard Behringer, Gigi Lozano) has produced a number of insights into the technology, including describing the mosaic nature of the resulting F0 animals, and establishing new methods to increase the rate of exact targeting (homology-driven repair). Collaborations have been established to develop and test novel ways to generate conditional mutations. New Cas enzymes with different sequence specificity and those providing more faithful activity will be tested. Our results will be widely shared with other transgenic facilities as they are validated. We are highly involved with both developing and with standardizing new technologies for use at MD Anderson and in transgenic facilities worldwide.