Genome Editing and Animal Models

项目总结/摘要 基因工程动物和细胞系被广泛用作模型，以确定潜在的机制 癌症病因和进展，并评估癌症治疗和预防的策略。基因组 编辑和动物模型共享资源（GEAM）已作为共享资源的大学 威斯康星州Carbone癌症中心（UWCCC）成立三十多年，是美国的精英机构之一。 各国提供生成、表征和保存编辑过的基因组所需的综合服务 动物模型GEAM的使命是为动物和细胞制造最先进的基因组编辑技术。 线以及UWCCC成员可以使用的冷冻保存服务。具体目标1： 新的、高度相关的、基因组编辑的或转基因的人类癌症动物模型。GEAM员工服务 UWCCC成员在实验规划的各个方面，包括设计高效的方法来实现 使用CRISPR、同源重组和基于转基因的基因组操作 方法;携带所需基因组编辑或转基因的动物的鉴定;以及最小化 偏离目标的编辑。我们在生殖生物学、胚胎操作和畜牧业方面的丰富经验 使GEAM能够编辑表现出低生殖能力的小鼠和大鼠品系的基因组。具体目标 2是开发克隆衍生的、基因组编辑的细胞系，用于癌症研究。在当前周期中， GEAM建立了一个管道，能够编辑各种已建立的细胞系的基因组， 安装了最先进的自动克隆跟踪系统具体目标3是提供最先进的 生殖生物学服务，使动物模型库，恢复和共享。GEAM精子 胚胎冷冻保存能力降低了动物饲养成本， 通过胚胎移植或体外受精。啮齿动物模型的重新推导消除了可能 危害研究或阻止动物模型输入到校园动物园或在合作者之间共享。 在本周期，GEAM继续通过购买新设备实现基础设施现代化 利用机构支持，采用最新最先进的基因组编辑方法， 训练有素的员工，以最好地满足我们的用户日益增长的技术挑战性需求。39个独特的 UWCCC计划成员在当前癌症中心支持补助金（CCSG）期间由GEAM提供服务 融资周期。CCSG的支持使GEAM的服务能够满足UWCCC的独特需求 成员，并以低于其他研究的商业供应商或类似核心的成本提供 高校GEAM的一流，成本效益，服务提高UWCCC调查人员进行 创新研究，推进中心的战略使命。来自UWCCC两年一次的调查数据 会员表示GEAM的服务是高品质的，对会员的研究至关重要。

PROJECT SUMMARY / ABSTRACT Genetically engineered animals and cell lines are widely used as models to define the mechanisms underlying cancer etiology and progression and to evaluate strategies for cancer treatment and prevention. The Genome Editing and Animal Models Shared Resource (GEAM) has served as a shared resource for the University of Wisconsin Carbone Cancer Center (UWCCC) for over three decades and is one of the elite facilities in the United States to offer the comprehensive services required to generate, characterize, and preserve genome edited animal models. GEAM’s mission is to make state-of-the-art genome editing technologies for animals and cell lines as well as cryopreservation services accessible to UWCCC members. Specific Aim 1 is to generate novel, highly relevant, genome edited or transgenic animal models of human cancers. GEAM staff serve UWCCC members in all aspects of experiment planning, including the design of efficient approaches to achieve the desired genome manipulation using CRISPR, homologous recombination, and transgene-based approaches; the identification of animals that carry the desired genome edit or transgene; and the minimization of off target edits. Our extensive experience in reproductive biology, embryo manipulation, and animal husbandry enable GEAM to edit the genomes of mouse and rat strains that exhibit low reproductive capacity. Specific Aim 2 is to develop clonally derived, genome edited, cell lines for use in cancer research. In the current cycle, GEAM established a pipeline that is capable of editing the genome of a wide variety of established cell lines and implemented a state-of-the-art automated clone tracking system. Specific Aim 3 is to provide state-of-the-art reproductive biology services that enable animal model banking, recovery, and sharing. GEAM’s sperm and embryo cryopreservation capabilities reduce animal husbandry costs and allow recovery of cancer models through embryo transfer or in vitro fertilization. Rederivation of rodent models eliminates pathogens that may compromise research or prevent animal model importation into campus vivaria or sharing between collaborators. In the current cycle, GEAM has continued to modernize our infrastructure through purchases of new equipment using institutional support, adopted the latest and most advanced genome editing methods, and added highly trained staff to best meet the increasing and technically challenging needs of our users. Thirty-nine unique UWCCC program members were served by GEAM during the current Cancer Center Support Grant (CCSG) funding cycle. Support from the CCSG enables GEAM’s services to be tailored to the unique needs of UWCCC members and provided at costs below those of commercial vendors or similar cores at other research universities. GEAM’s first rate, cost effective, services enhance the ability of UWCCC investigators to conduct innovative research and advance the strategic mission of the Center. Data from biennial surveys of UWCCC members indicate GEAM’s services are high quality and critical to our members’ research.