03 Genetically Engineered Mouse Facility

03 基因工程小鼠设施

• 批准号: 10212251
• 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/10212251
• 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.

项目总结：遗传标记小鼠工厂（GEMF） MD安德森基因工程小鼠设施（GEMF）提供了广泛的小鼠遗传学， 以合理的费用向调查员提供工程服务。提供的有偿服务包括转基因 小鼠生成（使用原核和胚泡注射），基因靶向（使用CRISPR/Cas9， 转座子介导的转基因，胚胎干细胞[ES]的电穿孔，和TARGATT 方案），小鼠胚胎和精子的存档，从研究者自己的小鼠产生ES细胞 线，以及鼠标线的重新推导。此外，咨询评估如何基因工程 动物可以为PI的项目做出贡献，并免费提供资助。一项服务， GEMF包括小鼠资源设施以提供试剂（例如，超数排卵激素 细胞培养基）、DNA质粒（产生用于基因靶向和原核表达的DNA构建体所需 注射）和Cre、LacZ、GFP和p53转基因小鼠（通常用于检测和/或产生 小鼠中的条件突变和用于探测肿瘤表型）。通过CCSG提供的支助 开发基金对于开发新的GEMF服务（sgRNA的体外转录）至关重要。的 该设施自1988年以来一直在使用，由Guillermina Lozano博士创办，他被任命为 2013年担任董事，目前由联席董事Jan Parker-Thornburg博士管理，他是 R50奖励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）， 其中342，039美元（44%）由CCSG资助。在目前的资助期内， 投资292，586美元用于资本设备，包括更换显微镜，自动注射 系统，和多功能电穿孔系统，可用于ES细胞和 CRISPR/Cas9胚胎电穿孔。所提供的服务在以下方面分配得相当平均： MD安德森的研究人员;代表所有16个CCSG项目的155名癌症中心成员使用了 在过去的6年里，GEMF服务。在第42年赠款中，同行审查的资金支持使用占 所有使用量的95%，要求CCSG资金支付44年总费用的41%（316，563美元）。的 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，导致超过201出版物，其中包括69（34%）的期刊与IF >10和146 (73%）。GEMF的具体目标是：目标1。来生产基因工程 MD安德森研究人员在癌症研究中使用两种方法进行研究所需的动物模型 传统和国家的最先进的技术，根据教师的需要。目标2.为MD提供服务 安德森研究人员推导和存档小鼠模型。目标3.使用制造商生产的鼠标 用于产生独特的ES细胞系的胚胎。目标4。为了提供专业知识和培训， 护理和处理GEM模型。