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内， 彭宁顿生物医学公司，目前为COBRE研究人员和其他教职员工生产小鼠 以及其他机构的研究人员。核心利用 原核显微注射和胚胎干细胞技术来控制基因表达， 小鼠TCF的目标是制造高质量的转基因和基因敲除小鼠 生产在技术上和资金上都很容易获得。TCF虽然不是正式的 在前几个资助周期中，由COBRE支持，一直在提供小鼠模型， 眼镜蛇科学家多年来。这些模型是COBRE的关键试剂 科学家在获得外部资金方面具有竞争力（表E1）。在下一个供资周期， COBRE将正式建立对转基因最先进方法的支持， 技术，以允许方法和服务的持续增长。这些新工具由 TCF将是非常有效的转化模型，用于临床前的基本证明， COBRE教师正在进行的概念研究。更具体地说，对于下一个授标周期， 我们也在为下一代的目标定位战略做准备， 不需要胚胎干细胞。使用工程化核酸酶的靶向基因组编辑具有 在很大程度上是由集群的出现，定期间隔，短回文 重复序列（CRISPR）技术，一种重要的新方法，用于产生RNA引导的 核酸酶，如Cas9，具有可定制的特异性。基因组编辑介导的这些 核酸酶可用于在广泛的生物学领域中快速、容易和有效地修饰内源基因， 各种细胞类型和传统上难以操纵的生物体 遗传的这项技术有可能消除费时费力的 为小鼠设计靶向结构，但更重要的是为基因工程打开了大门。 几乎所有物种的目标。这一点很重要，因为许多COBRE教师使用 大鼠作为其优选的临床前模型。我们的计划是开始在老鼠身上测试这项技术 然后用老鼠作为基因操作的模型。TCF的具体目标 1）利用转基因和基因打靶技术建立小鼠模型， 模拟人类疾病状态，如肥胖、胰岛素抵抗和脂质失调 2）根据需求寻求新的CRISPR方法和靶向策略 来自COBRE教师和试点和可行性资金的接受者。