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

描述(申请人提供)：神经生物学的一个主要目标是确定功能神经通路。经典的解剖追踪技术可以确定大脑区域之间的连通性。损伤研究和解剖学上受限的药物输注有助于在大体水平上确定这些脑区的功能作用。在过去的15年里，小鼠遗传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的分子机制的能力。然而，将遗传学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复杂的大脑神经解剖结构上的能力。目前应用的目标是开发一系列48个驱动系，用于产生解剖学上受限和可诱导的基因敲除。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，最广泛使用的解剖受限和时间依赖的基因功能操纵工具分别是Cre重组酶和TTA反式激活因子，但神经科学界可用的驱动因素很少。此外，大多数CRE驱动系不允许严格的时间控制，例如敲除成人组织中的基因的能力。我们计划使用转基因方法来表达Cre和TTA，由所选的24个基因的调控元件驱动。默认的方法是通过ES细胞中的同源重组来靶向；也将利用原核注射的载体(包括BAC)。驱动线将允许我们实现对神经系统不同区域中重组酶活性的时间和空间控制。将通过监控电池类型和依赖时间的CRE活动来验证驱动程序线。行为学研究将确保转基因不会影响神经功能。24个驾驶员基因座的选择代表了参与这一应用的四名研究人员的专业知识，并与神经科学界相关，涉及感觉生物学和疼痛、CMS和干细胞发育、边缘系统以及学习和记忆。从临床的角度来看，产生的线条应该与神经精神障碍和成瘾的研究特别相关(例如。以多巴胺系统为靶点)和神经退行性疾病(例如以边缘系统和成人干细胞为靶点)。

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).