Transgenics Core

转基因核心

• 批准号: 9978081
• 负责人: Randall Lee Mynatt
• 金额: $ 12.89万
• 依托单位: LSU PENNINGTON BIOMEDICAL RESEARCH CTR
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-01 至 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9978081
• 关键词: Address; Alleles; Award; CRISPR/Cas technology; Centers of Research Excellence; Comparative Biology; Consultations; Consumption; Core Facility; DNA; Development; Diabetes Mellitus; Engineering; Faculty; Funding; Gene Expression; Gene Targeting; Generations; Genes; Genetic Models; Genetically Modified Animals; Goals; Growth; Guide RNA; Institution; Insulin Resistance; Knockout Mice; Louisiana; Mediating; Mentors; Metabolic Diseases; Methods; Microinjections; Mission; Modeling; Monitor; Mus; Obesity; Organism; Pre-Clinical Model; Procedures; Production; Productivity; Rattus; Reagent; Research; Research Personnel; Scientist; Services; Specificity; Techniques; Technology; Testing; Time; Tissues; Training; Transgenic Organisms; Work; base; cell type; embryonic stem cell; engineered nucleases; genetic manipulation; genome editing; human disease; innovation; knockout gene; lipid metabolism; mouse model; next generation; novel strategies; nuclease; pre-clinical; promoter; stem cell technology; tool; translational model; virtual

The Transgenic Core Facility (TCF) is located within the Comparative Biology Facility at Pennington Biomedical and currently produces mice for COBRE investigators and other faculty at Pennington Biomedical as well as investigators at other institutions. The core utilizes pronuclear microinjection and embryonic stem cell technologies to control gene expression in mice. The objective of the TCF is to make high quality transgenic and gene knockout mouse production readily accessible, both technically and financially. The TCF, although not formally supported by the COBRE in the previous funding cycles, has been providing mouse models to COBRE scientists for several years. These models have been crucial reagents to allow COBRE scientists to be competitive in obtaining external funding (Table E1). For this next funding cycle, the COBRE will formally establish support for state-of-the-art methods in transgenic technologies to allow continued growth of methods and services. These new tools produced by the TCF will be highly effective translational models for the essential pre-clinical proof of concept studies being conducted by COBRE faculty. More specifically for the next award cycle, we are also preparing for what appears to be the next generation of targeting strategies that do not require embryonic stem cells. Targeted genome editing using engineered nucleases has been largely fueled by the emergence of clustered, regularly interspaced, short palindromic repeat (CRISPR) technology, an important new approach for generating RNA-guided nucleases, such as Cas9, with customizable specificities. Genome editing mediated by these nucleases can be used to rapidly, easily and efficiently modify endogenous genes in a wide variety of cell types and in organisms that have traditionally been challenging to manipulate genetically. This technology has the potential to eliminate the laborious and time-consuming engineering of targeting constructs for mice, but more importantly opens the doors for gene targeting in virtually any species. This is significant because a number of COBRE faculty use rats as their preferred pre-clinical model. Our plan is to begin testing this technology in mice and then progress to rats as a model for genetic manipulation. The Specific Aims of the TCF are to: 1) To utilize transgenic and gene targeting techniques to generate mouse models that mimic human disease states, such as obesity, insulin resistance, and dysregulation of lipid metabolism, and 2) Pursue new CRISPR methods and targeting strategies based on the needs of from COBRE faculty and recipients of Pilot and Feasibility funding.

转基因核心设施（TCF）位于比较生物学设施中 Pennington生物医学，目前为毛毛调查员和其他教职员工生产小鼠 在彭宁顿生物医学以及其他机构的研究人员。核心使用 前核微注射和胚胎干细胞技术控制基因表达 老鼠。 TCF的目的是使高质量的转基因和基因敲除鼠标 在技​​术上和财务上都可以轻松访问生产。 TCF，尽管不是正式的 在以前的资金周期中的毛绒支持下，一直在提供鼠标模型 毛科学家已经有好几年了。这些模型一直是允许毛病的关键试剂 科学家在获得外部资金方面具有竞争力（表E1）。对于下一个资金周期， 该鞋底将正式建立对转基因最新方法的支持 允许持续增长方法和服务的技术。这些新工具由 TCF将是高效的转化模型，用于基本的临床前证明 概念研究由Cobre教师进行。专门针对下一个奖励周期， 我们还在准备似乎是下一代的目标策略 不需要胚胎干细胞。使用工程核酸酶的靶向基因组编辑已有 在很大程度上被聚集，定期插入的，短的alindromic的出现促成 重复（CRISPR）技术，这是生成RNA引导的重要新方法 具有可自定义特异性的核酸酶，例如Cas9。这些介导的基因组编辑 核酸酶可用于快速，易于有效地修饰宽阔的内源基因 传统上挑战的各种细胞类型和生物体中 遗传。这项技术有可能消除费力且耗时的 小鼠靶向构建的工程，但更重要的是为基因打开门 靶向几乎任何物种。这很重要，因为许多毛茸茸的教师使用 大鼠是他们首选的临床前模型。我们的计划是开始在小鼠中测试这项技术 然后发展为大鼠作为基因操纵的模型。 TCF的具体目的 是：1）利用转基因和基因靶向技术来生成鼠标模型 模仿人类疾病状态，例如肥胖，胰岛素抵抗和脂质失调 新陈代谢，以及2）根据需求追求新的CRISPR方法和针对策略 来自Cobre的教职员工和飞行员和可行性资金的接受者。