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识别功能性神经元通路。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药理学药物的输注有助于在大体水平上确定这些脑区域的功能作用。在过去的15年里，老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复杂的大脑神经解剖学上的能力。当前应用的目标是开发一系列48个驱动系，用于产生解剖学上受限的和可诱导的基因敲除。为了最大的遗传效用神经科学界菌株将开发在纯C57BL/6背景。在解剖受限和时间依赖的小鼠基因功能操作中，最广泛使用的工具分别是cre -重组酶和ta -反激活因子，但很少有驱动线可供神经科学界使用。此外，大多数cre驱动系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。我们计划用转基因的方法表达由24个选定基因的调控元件驱动的CRE和tTA。默认的方法是通过胚胎干细胞的同源重组靶向；还将利用原核注射（包括BACs）的载体。驱动线将使我们能够在神经系统的不同区域实现重组酶活性的时间和空间控制。驱动线将通过监测细胞类型和时间相关的CRE活动来验证。行为学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程序的四位研究者的专业知识，并且与神经科学社区相关，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，生成的谱线应该与神经精神疾病和成瘾的研究特别相关。靶向多巴胺系统)和神经退行性疾病（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).