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

描述（由申请人提供）：神经生物学的一个主要目标是鉴定功能性神经元通路。经典的解剖追踪技术允许确定大脑区域之间的连接。病变研究和解剖学上限制的药物注入有助于在总体水平上确定这些脑区的功能作用。在过去的15年中，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测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学变化映射到大脑复杂神经解剖学上的能力。本申请的目标是开发一系列48个驱动系，用于产生解剖学限制性和诱导性基因敲除。为了对神经科学界最大的遗传效用，将在纯C57 BL/6背景下开发菌株。最广泛使用的工具，在小鼠中的基因功能的解剖学上的限制和时间依赖性的操作是CRE重组酶和tTA-反式激活因子，分别，但很少的驱动程序线可用于神经科学界。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的方法来表达CRE和tTA驱动的24个选定的基因的调控元件。默认方法是通过ES细胞中的同源重组进行靶向;也将使用原核注射载体（包括BAC）。驱动线将允许我们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。将通过监测细胞类型和时间依赖性CRE活性来验证驱动细胞系。行为研究将确保转基因不会影响神经功能。24个驱动基因座的选择代表了参与该应用的四名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。从临床的角度来看，所产生的线条应该特别与神经精神疾病和成瘾的研究相关（例如，靶向多巴胺系统）和神经变性疾病（例如靶向边缘系统和成体干细胞）。

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).