Developing drivers for neuron type-specific gene expression

开发神经元类型特异性基因表达的驱动程序

• 批准号: 8935927
• 负责人: Oliver Hobert
• 金额: $ 62.6万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-26 至 2017-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8935927
• 关键词: Acetylcholine; Adult; Aminobutyric Acids; Binding Sites; Brain; Caenorhabditis elegans; Cell Count; Cells; DNA; Dissection; Elements; Enzymes; Future; Gene Expression; Generations; Genes; Genetic; Genomics; Glutamates; Health; Individual; Integrase; LacZ Genes; Maps; Mediating; Molecular; Monitor; Mus; Mutate; Nature; Nematoda; Nervous system structure; Neuraxis; Neurons; Neurosciences; Neurotransmitters; Nucleic Acid Regulatory Sequences; Pattern; Phenotype; Population; Positioning Attribute; Proteins; Regulatory Element; Reporter; Specificity; System; Testing; Transgenic Mice; Transgenic Organisms; Work; acetylcholine transporter; base; cell type; cholinergic; gene function; genetic manipulation; insight; interest; knockout gene; mouse genome; neuronal circuitry; novel; optogenetics; theories; tool; transcription factor

 DESCRIPTION (provided by applicant): Driver lines that direct Cre protein to specific neuron types have proven to be invaluable tools to not only visualize specific neuron types but also to manipulate their activity through the Cre- mediated activation of optogenetic probes or to assess gene function by Cre-mediated gene knockout. Most Cre driver lines, such as BAC-based Cre drivers or knock-ins of Cre into specific loci, monitor the complete expression pattern of entire genetic loci. However, very few genes are exclusively expressed in very small populations of specific neuron types and this lack of cellular specificity limits the use of these driver lines. W propose here to develop transgenic mouse driver lines that direct Cre expression to very restricted numbers of neuronal cell types in different regions of the mouse brain, thereby providing tools to precisely map their function and molecular composition. To achieve this aim, we aim to test the hypothesis - built from our past work in the nematode C.elegans - that the cis-regulatory control elements of the mouse loci that encode the vesicular transporters for the four main neurotransmitter systems in the vertebrate central nervous system, glutamate and -aminobutyric acid (GABA) and acetylcholine (ACh) are composed of a modular assembly of individual, highly cell type-specific cis-regulatory elements. We will experimentally test the hypothesis that the expression of individual, isolated cis-regulatory elements may subdivide cholinergic, glutamatergic and GABAergic domains into restricted and perhaps novel domains of the mouse central nervous system and thereby constitute reproducible and highly specific drivers for directing the expression of genes that allow the genetic manipulation of neurons and neuronal circuits. This cis-regulatory dissection approach may solve the specificity problem of most currently available driver lines that are unable to exclusively target restricted numbers of cells.

 描述（由适用提供）：将CRE蛋白转向特定神经元类型的驱动线已被证明是宝贵的工具，不仅可以可视化特定的神经元类型，还可以通过介导的光遗传学问题激活或通过CRE介导的基因敲除来评估基因功能来操纵其活性。大多数CRE驱动线（例如基于BAC的CRE驱动器或Cre inscre ins to to特定局部）监测整个遗传基因座的完整表达模式。但是，很少有基因在非常小的特定神经元类型中仅表达，并且缺乏细胞特异性限制了这些驱动线的使用。 w此处的提案要开发转基因小鼠驱动线，将CRE表达引导至小鼠大脑不同区域中非常有限的神经元细胞类型，从而提供了精确绘制其功能和分子组成的工具。为了实现这一目标，我们旨在测试假设 - 根据我们过去在线虫C.Elegans中的工作构建的 - 鼠标基因座的顺式调节控制元素编码四个主要神经递质系统的囊泡转运蛋白，用于脊椎动物中枢神经系统，谷氨酸和氨基酸（gaba）和抗氨基酸含量（gaba）和actyylcy offerty offecrine and Actylchol offectional of。个体，高度细胞类型特异性的顺式调节元件。我们将在实验上检验以下假设：个体，孤立的顺式调节元件的表达可能会亚细胞，谷氨酸能和GABA能结构域对小鼠中央神经系统的受限和可能的新领域进行促胆碱能，从而构成neurons of Neuronon和Neurononon的表达，从而构成了基因和高度特定的驱动因素。这种顺式调节方法可能会解决无法仅针对限制数量的细胞数量的大多数当前可用驾驶员线的特异性问题。