C57BI/6 Mouse Lines Expressing CRE-Recombinase in the Nervous System

在神经系统中表达 CRE 重组酶的 C57BI/6 小鼠系

• 批准号: 7676891
• 负责人: Ulrich Mueller
• 金额: $ 160.72万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-08 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7676891
• 关键词: Adult; Affect; Alleles; Applications Grants; Behavioral; Biological Models; Biological Process; Biology; Brain; Brain region; Cells; Clinical; Communities; Complex; Data; Development; Disease; Ensure; Enterobacteria phage P1 Cre recombinase; Excision; Exons; Gene Expression; Generations; Genes; Genetic; Genetic Recombination; Genomics; Goals; Infusion procedures; Injection of therapeutic agent; Introns; Knock-in Mouse; Knock-out; Laboratories; Learning; Lesion; Limbic System; Maps; Memory; Molecular Genetics; Molecular Probes; Monitor; Mus; Mutation; Nervous system structure; Neuroanatomy; Neurobiology; Neurodegenerative Disorders; Neurons; Neurosciences; Pain; Pathway interactions; Pattern; Polyadenylation; Regulatory Element; Relative (related person); Research; Research Personnel; Role; Sensory; Series; Signal Transduction; Simian virus 40; Site; Smell Perception; Stem Cell Development; System; Techniques; Time; Tissues; Trans-Activators; Transgenes; Transgenic Mice; Transgenic Organisms; Upper arm; addiction; adult stem cell; base; cell type; design and construction; dopamine system; embryonic stem cell; experience; gene function; genetic manipulation; homologous recombination; in vivo; inhibitor/antagonist; interest; neuropsychiatry; premature; prevent; programs; recombinase; technique development; tool; vector

DESCRIPTION (Provided by Applicant): A major goal in neurobiology is identifying functional neuronal pathways. Classical anatomical tracing techniques allow connectivity between brain regions to be determined. Lesion studies and anatomically restricted infusion of pharmacological agents have helped to identify the functional role of these brain regions at a gross level. The development of techniques for the genetic manipulation of the mouse during the past 15 years has greatly expanded our ability to probe the molecular mechanisms of biological function in a mammalian model system. However, one of the difficulties in the application of the genetic approach to the nervous system is the relative lack of ability to map molecular genetic changes onto the complex neuroanatomy of the brain. The goal of the current application is to develop a series of 48 driver lines for the generation of anatomically restricted and inducible gene knock-outs. For greatest genetic utility to the neuroscience community the strains will be developed on a pure C57BL/6 background. The most widely used tools for anatomically restricted and time dependent manipulation of gene function in the mouse are the CRE-recombinase and the tTA-transactivator, respectively, but very few driver lines are available to the neuroscience community. Furthermore, most CRE-driver lines do not allow strict temporal control, such as the ability to knock-out genes in adult tissues. We plan to use transgenic approaches to express CRE and tTA driven by the regulatory elements of 24 chosen genes. The default approach is targeting via homologous recombination in ES cells; vectors for pronuclear injections (including BACs) will also be utilized. The driver lines will allow us to achieve temporal and spatial control of recombinase activity in disparate regions of the nervous system. Driver lines will be validated by monitoring cell type and time dependent CRE activity. Behavioral studies will ensure that the transgenes do not affect neuronal function. The choice of the 24 driver loci represents the expertise of the four investigators involved in this application and are of relevance for the neuroscience community, covering sensory biology and pain, CMS and stem cell development, the limbic system, and learning and memory. From a clinical perspective the lines generated should be particularly relevant to the study of Neuropsychiatric disorders and addiction (eg. targeting the dopamine system) and neurodegenerative disorders (e.g. targeting the limbic system and adult stem cells).

描述（由申请人提供）：神经生物学的主要目标是识别功能性神经元途径。经典的解剖示踪技术允许确定大脑区域之间的连通性。病变研究和解剖学限制药理学剂的注入有助于确定这些大脑区域在总水平上的功能作用。在过去15年中，用于小鼠基因操纵的技术的发展大大扩展了我们在哺乳动物模型系统中探测生物功能分子机制的能力。但是，在神经系统中应用遗传方法的困难之一是将分子遗传变化映射到大脑复杂的神经解剖学的能力相对缺乏能力。当前应用的目的是开发一系列48个驱动线，以生成解剖学上受限制和诱导的基因敲除。对于神经科学界的最大遗传效用，应在纯C57BL/6背景下开发菌株。用于解剖学上的最广泛使用的工具和对小鼠基因功能的时间依赖性操纵分别是CRE-异构酶和TTA转移剂，但神经科学社区的驱动线很少。此外，大多数CRE驱动器线不允许严格的时间控制，例如在成年组织中敲除基因的能力。我们计划使用转基因方法表达由24个基因的调节元素驱动的CRE和TTA。默认方法是通过ES细胞中的同源重组来靶向。还将利用前核注射载体（包括BAC）。驱动线将使我们能够在神经系统的不同区域中实现重组酶活性的时间和空间控制。驱动线将通过监视单元格类型和时间依赖的CRE活动来验证。行为研究将确保转基因不会影响神经元功能。 24个驱动程序基因座的选择代表了参与此应用的四名研究人员的专业知识，并且与神经科学社区相关，涵盖了感觉生物学和疼痛，CMS和干细胞开发，边缘系统以及学习和记忆。从临床角度来看，产生的线应与神经精神疾病和成瘾的研究特别相关（例如，靶向多巴胺系统）和神经退行性疾病（例如，靶向缘缘系统和成年干细胞）。