Transgenic and Targeted Mutagenesis Laboratory Shared Resource

摘要-转基因和靶向诱变实验室 转基因和靶向突变实验室(TTML)是西北大学范围内的资源 致力于基因工程小鼠的生成、导入、恢复/再衍生和冷冻保存。 TTML为Lurie癌症中心(LCC)成员提供广泛的服务，包括生成 通过显微注射转基因、基因编辑和靶向转基因小鼠模型；基因打靶/编辑 胚胎干细胞(ES细胞)；胚胎干细胞显微注射到囊胚中以产生生殖系能力的嵌合体； 从种质中引进和回收/再衍生小鼠品系；小鼠精子的冷冻保存 和胚胎。自上次审查以来，该核心已经引入了全面的、从头到尾的突变能力。 这些变化对于促进新兴基因组工程的发展是必要的。 CRISPR等技术。TTML团队与LCC成员合作设计定制的基因 修改策略和基因分型分析，并向调查人员提供确认的创始人。这些新的 除了我们创建转基因和ESC衍生突变系的传统方法之外，我们的能力还能够 研究人员有机会使用主要的方法通过 设施。 TTML设施对于Lurie癌症中心研究人员的研究项目至关重要，他们使用 小鼠模型，用于研究恶性肿瘤的潜在机制，并验证翻译项目的靶点。它 提供必要的基础设施，使中心的研究人员能够获得转基因技术 通常需要昂贵的微注射设备和具有微注射专业知识的熟练工作人员， 显微外科、胚胎操作、畜牧业、胚胎干细胞培养和基因组工程。此外, TTML的工作人员为所有转基因相关技术提供内部咨询和经常性指导，包括 CRISPR介导的基因组编辑系统、转基因和靶向载体设计、适当筛选 方正小鼠的策略、DNA提纯方法、选育与分析、适宜的引进方法 小鼠品系和冷冻保存选项。由于将TTML作为资源包含在Lurie癌症中 中心成立于1995年，中心调查人员一直是使用TTML的NU教职员工的主要群体， 强调其在LCC总体研究任务中的关键作用。

ABSTRACT – TRANSGENIC AND TARGETED MUTAGENESIS LABORATORY The Transgenic and Targeted Mutagenesis Laboratory (TTML) is a Northwestern University-wide resource dedicated to the generation, import, recovery/rederivation, and cryopreservation of genetically engineered mice. TTML provides Lurie Cancer Center (LCC) members a broad range of services including generation of transgenic, gene edited, and targeted transgenic mouse models via microinjection; gene targeting/editing of embryonic stem (ES) cells; ES cell microinjection into blastocysts to create germline competent chimera; importation and recovery/rederivation of mouse lines from germplasm; and cryopreservation of mouse sperm and embryos. Since the last review, the core has introduced full-scale, start-to-finish mutagenesis capabilities. These changes were necessary to facilitate the development of newly emerging genome engineering technologies such as CRISPR. The TTML team works with LCC members to design customized genetic modification strategies and genotyping assays, and provides confirmed founders to investigators. These new capabilities, in addition to our traditional methods for creating transgenic and ESC derived mutant lines, enable investigators the opportunity to employ overarching approaches to genetic modification mutagenesis through the facility. The TTML facility is essential for the research programs of investigators of the Lurie Cancer Center who use mouse models to study mechanisms underlying malignancy and validate targets for translational projects. It provides the necessary infrastructure that allows the Center’s investigators access to transgenic technology that normally requires expensive microinjection equipment and skilled staff with expertise in microinjection, microsurgeries, embryo manipulation, animal husbandry, ES cell culture, and genome engineering. In addition, TTML staff provide in-house consultation and recurrent guidance on all transgenic-related technologies including CRISPR-mediated genome editing systems, transgenic and targeting vector design, appropriate screening strategies, DNA purification methods, breeding and analysis of founder mice, appropriate methods for importing mouse lines, and cryopreservation options. Since the inclusion of the TTML as a resource within the Lurie Cancer Center in 1995, Center investigators have consistently been the primary group of NU faculty utilizing the TTML, emphasizing its pivotal role in the overall research mission of the LCC.