03 Genetically Engineered Mouse Facility

03 基因工程小鼠设施

• 批准号: 10466981
• 负责人: Jan Parker-Thornburg
• 金额: $ 48.81万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-08-28 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10466981
• 关键词: Address; American; Animal Model; Animals; Archives; Area; Award; Breeding; Budgets; CRISPR/Cas technology; Cancer Center; Cancer Center Support Grant; Capital Financing; Caring; Clustered Regularly Interspaced Short Palindromic Repeats; Communities; Consultations; Cryopreservation; DNA; Derivation procedure; Detection; Development; E-learning; ES Cell Line; Educational process of instructing; Educational workshop; Electroporation; Embryo; Equipment; Faculty; Fertilization in Vitro; Funding; Gene Targeting; Gene Transfer Techniques; Gene-Modified; Generations; Genes; Genetic; Genetic Engineering; Genetic Transcription; Genetically Engineered Mouse; Grant; Hormones; Human Resources; In Vitro; Injections; Institution; Japan; Journals; Knock-in; Knock-out; Knowledge; Laboratory Animal Science; LacZ Genes; Libraries; Link; Mediating; Methodology; Microscope; Mission; Modeling; Mus; Mutation; Oocytes; Paper; Peer Review; Phenotype; Procedures; Protocols documentation; Publications; Publishing; Reagent; Reproducibility; Research; Research Personnel; Resource Development; Resources; Science; Services; Stem Cell Development; Superovulation; System; TP53 gene; Techniques; Testing; Texas; Training; Transgenic Mice; Transgenic Model; Transgenic Organisms; Update; animal care; animal facility; animal resource; anticancer research; base; blastocyst; conditional mutant; cost; embryonic stem cell; experimental study; in vivo; innovation; member; mouse genetics; mouse model; new technology; plasmid DNA; programs; sperm cell; tumor; webinar

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.

项目摘要：基因工程小鼠设施 (GEMF) MD 安德森基因工程小鼠设施 (GEMF) 提供广泛的小鼠遗传 以合理的成本向调查人员提供工程服务。提供的有偿服务包括转基因 小鼠生成（使用原核和囊胚注射）、基因靶向（使用 CRISPR/Cas9、 转座子介导的转基因、胚胎干 [ES] 细胞的电穿孔和 TARGATT@ 方案），小鼠胚胎和精子的存档，从研究人员自己的小鼠中生成 ES 细胞 线，以及鼠标线的重新衍生。此外，还提供有关评估基因工程如何进行的咨询 动物可以为 PI 的项目做出贡献，并且免费提供赠款支持。独有的服务 GEMF 包括小鼠资源设施，以提供试剂（例如超排卵激素、ES 细胞培养基）、DNA 质粒（生成用于基因靶向和原核的 DNA 构建体所需） 注射），以及 Cre、LacZ、GFP 和 p53 转基因小鼠（通常用于检测和/或生成 小鼠条件突变和探测肿瘤表型）。通过 CCSG 提供的支持 开发基金对于开发新的 sgRNA 体外转录 GEMF 服务至关重要。这 该设施自 1988 年起投入使用，由 Guillermina Lozano 博士创办，并被任命为 2013 年担任董事，目前由联合董事 Jan Parker-Thornburg 博士管理，他是 R50 奖。该设施的年度预算为 775,887 美元（其中 74% 用于高级专家人员的费用）， 其中 342,039 美元 (44%) 由 CCSG 资金支持。在本资助期内，该机构 投资 292,586 美元购买资本设备，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，又可用于 CRISPR/Cas9 胚胎电穿孔。所提供的服务分布相当均匀 MD 安德森的研究人员；代表所有 16 个 CCSG 项目的 155 名癌症中心成员已使用 过去 6 年的 GEMF 服务。在第 42 年拨款中，同行评审的资金支持用途占 95% 的使用量和 CCSG 资金被要求支付 44 年总支出的 41%（316,563 美元）。这 GEMF 生产的动物和材料为马里兰大学的高影响力科学做出了重大贡献 Anderson 发表了超过 201 篇论文，其中 69 篇 (34%) 发表在 IF >10 和 146 的期刊上 (73%) 在 IF >5 的期刊中。 GEMF 的具体目标是： 目标 1. 生产基因工程产品 MD 安德森研究人员在癌症研究中需要使用两种动物模型 根据教师的需要，采用传统和最先进的技术。目标 2. 为 MD 提供服务 安德森研究人员导出并存档小鼠模型。目标 3. 使用研究者生产的鼠标 胚胎用于产生独特的 ES 细胞系。目标 4. 为一代人提供专业知识和培训， GEM 模型的保养和处理。