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: 7936245
• 负责人: Ulrich Mueller
• 金额: $ 161.63万
• 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 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7936245
• 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; addiction; adult stem cell; arm; 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）：神经生物学的一个主要目标是识别功能性神经元通路。经典的解剖追踪技术可以确定大脑区域之间的连接性。病变研究和解剖学上限制的药物输注有助于在总体水平上确定这些大脑区域的功能作用。过去15年里，小鼠基因操作技术的发展极大地扩展了我们在哺乳动物模型系统中探索生物功能分子机制的能力。然而，将遗传方法应用于神经系统的困难之一是相对缺乏将分子遗传变化映射到大脑复杂的神经解剖学上的能力。当前应用的目标是开发一系列 48 个驱动线，用于生成解剖学限制和诱导基因敲除。为了给神经科学界带来最大的遗传效用，这些菌株将在纯 C57BL/6 背景下开发。在小鼠中对基因功能进行解剖学限制和时间依赖性操作的最广泛使用的工具分别是 CRE 重组酶和 tTA 反式激活子，但神经科学界可用的驱动线很少。此外，大多数 CRE 驱动系不允许严格的时间控制，例如敲除成体组织中基因的能力。我们计划使用转基因方法来表达由 24 个选定基因的调控元件驱动的 CRE 和 tTA。默认方法是通过 ES 细胞中的同源重组进行靶向；还将利用用于原核注射的载体（包括 BAC）。驱动线将使我们能够实现神经系统不同区域重组酶活性的时间和空间控制。驱动细胞系将通过监测细胞类型和时间依赖性 CRE 活性进行验证。行为研究将确保转基因不会影响神经元功能。 24 个驱动基因座的选择代表了参与该应用的四位研究人员的专业知识，并且与神经科学界相关，涵盖感觉生物学和疼痛、CMS 和干细胞发育、边缘系统以及学习和记忆。从临床角度来看，生成的细胞系应该与神经精神疾病和成瘾（例如针对多巴胺系统）和神经退行性疾病（例如针对边缘系统和成体干细胞）的研究特别相关。