03 Genetically Engineered Mouse Facility

项目摘要：基因工程小鼠设施(GEMF) MD Anderson基因工程小鼠设施(GEMF)提供了广泛的小鼠基因 以合理的成本为调查人员提供工程服务。提供的有偿服务包括转基因 小鼠的产生(使用原核和囊胚注射)，基因打靶(使用CRISPR/Cas9， 转座子介导的转基因、胚胎干细胞的电穿孔和TARGATT@ )，小鼠胚胎和精子的存档，从研究人员自己的小鼠中产生ES细胞 线条和鼠标线条的重新派生。此外，为评估如何利用基因工程提供咨询 动物可以为公益机构的项目做出贡献，并免费提供赠款支持。一项独一无二的服务 GEMF是包括老鼠资源设施以提供试剂(例如，超排激素，ES 细胞培养液)、DNA质粒(产生用于基因打靶和原核的DNA构建所必需的 注射)和Cre、LacZ、GFP和P53转基因小鼠(通常用于检测和/或世代 小鼠的条件性突变和探测肿瘤表型)。通过CCSG提供的支持 开发资金对于开发新的GEMF服务--体外转录sgRNA至关重要。这个 该设施自1988年开始服务，由Guillermina Lozano博士创办，他被任命为 2013年担任主任，目前由联合主任简·帕克-索恩伯格博士管理，他是 R50大奖。该设施的年度预算为775,887美元(74%用于高级专家人员的费用)， 342,039美元(44%)，其中CCSG资金支持。在本授权期内，该机构 为资本设备投资292,586美元，包括更换显微镜、自动注射 系统，和多功能电穿孔系统，既可用于ES细胞，也可用于 CRISPR/Cas9胚胎电穿孔。提供的服务相当平均地分布在 MD Anderson的调查人员；代表CCSG所有16个项目的155名癌症中心成员使用了 过去6年的GEMF服务。在赠款Yr42中，同行评审的资金支助使用情况占了 所有使用量的95%，CCSG资金被要求支付44财年总费用的41%(316,563美元)。这个 GEMF生产的动物和材料为MD的高影响力科学做出了重大贡献 安德森说，已经发表了201多篇论文，其中69篇(34%)发表在IF&>10和146的期刊上 (73%)发表在IF&GT；期刊上5.GEMF的具体目标是：目标1.生产转基因产品 MD Anderson研究人员在癌症研究中使用这两种方法所需的动物模型 传统的和最新的技术，基于教师的需要。目标2.为MD提供服务 安德森研究人员对小鼠模型进行了推导和存档。目标3.使用研究人员生产的小鼠 胚胎用于产生独特的ES细胞系。目标4.为这一代人提供专门知识和培训， 宝石模型的保养和搬运。

PROJECT SUMMARY: GENETICALLY ENGINEERED MOUSE FACILITY (GEMF) The MD Anderson Genetically Engineered Mouse Facility (GEMF) provides a wide range of mouse genetic engineering services to investigators at a reasonable cost. Compensated services provided include transgenic mouse generation (using pronuclear and blastocyst injection), gene targeting (using CRISPR/Cas9, transposon-mediated transgenesis, the electroporation of embryonic stem [ES] cells, and TARGATT@ protocols), archiving of mouse embryos and sperm, generation of ES cells from investigators' own mouse lines, and rederivation of mouse lines. In addition, consultation for assessing how genetically engineered animals can contribute to PIs' projects and grant support are provided at no cost. A service unique to the GEMF is the inclusion of a Mouse Resource Facility to provide reagents (e.g., superovulation hormones, ES cell media), DNA plasmids (required for generating DNA constructs for gene targeting and pronuclear injection), and Cre, LacZ, GFP, and p53 transgenic mice (commonly used for the detection and/or generation of conditional mutations in mice and for probing tumor phenotypes). Support provided through CCSG Development Funds was essential for developing the new GEMF service of in vitro transcription of sgRNA. The facility has been in service since 1988 and was started by Dr. Guillermina Lozano, who was appointed director in 2013, and is currently managed by Dr. Jan Parker-Thornburg, the co-director, who is a recipient of an R50 award. The facility has an annual budget of $775,887 (74% for costs for highly expert personnel), $342,039 (44%) of which is supported with CCSG funding. During the present grant period, the institution invested $292,586 in funding for capital equipment, including replacement microscopes, automated injection systems, and a multi-functional electroporation system that can be used both for ES cells and for CRISPR/Cas9 embryo electroporation. Services provided have been fairly evenly distributed among investigators at MD Anderson; 155 cancer center members representing all 16 CCSG programs have used GEMF services over the past 6 years. In grant Yr42, peer-reviewed funding–supported usage accounted for 95% of all usage, and CCSG funds are requested to cover 41% of total expenses in Yr44 ($316,563). The animals and materials produced by the GEMF have contributed significantly to high-impact science at MD Anderson that has resulted in more than 201 publications, including 69 (34%) in journals with IF >10 and 146 (73%) in journals with IF >5. The GEMF's specific aims are: Aim 1. To produce the genetically engineered animal models required by MD Anderson investigators for their studies in cancer research using both traditional and state-of-the-art techniques, based on the need of the faculty. Aim 2. To provide services to MD Anderson investigators to derive and archive mouse models. Aim 3. To use investigator-produced mouse embryos for the generation of unique ES cell lines. Aim 4. To provide expertise and training in the generation, care, and handling of GEM models.