General method for targeting genes to specific neuronal subtypes in mammals

将基因靶向哺乳动物特定神经元亚型的通用方法

基本信息

批准号：
8529011
负责人：
ANTHONY M ZADOR
金额：
$ 23.63万
依托单位：
COLD SPRING HARBOR LABORATORY
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-04-01 至 2015-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8529011
关键词：
Animal Model Animals Base Pairing Behavior Behavioral Paradigm Biological Models Brain Brain region Catalytic RNA Cells Coupled DRD2 gene Disease Dopamine Dopamine Receptor Gene Targeting Generations Genes Genetic Goals Halorhodopsins Human Investments Knock-in Mouse Learning Maintenance Mammals Messenger RNA Methods Molecular Monkeys Mus Neurons Organism Play Preparation Primates Psychological reinforcement RNA RNA Splicing Rattus Reagent Reporter Genes Research Rodent Role Specificity Therapeutic Studies Trans-Splicing Transcript Transgenes Transgenic Mice Transgenic Organisms Viral Virus Work addiction base cell type fluorophore genetic manipulation human subject neural circuit neuropsychiatry novel novel strategies optogenetics public health relevance receptor recombinant virus recombinase selective expression tool transgene expression

项目摘要

DESCRIPTION (provided by applicant): The goal of this project is to develop a broadly applicable method for convenient cell-type specific expression of transgenes, such as optogenetic reagents, in rat, primate and other mammalian brains. Circuits in the brain consist of diverse neuronal subtypes which are often defined by the selective expression of specific genes. For example, neurons defined by the expression of the D1R and D2R dopamine receptors represent distinct neuronal subpopulations which play distinct roles in reinforcement learning and addiction. The ability to target the expression of a growing number of molecular tools, including flourophores and optogenetic reagents, to such genetically-defined neuronal subtypes provides a powerful method for dissecting neural circuits. However, current approaches for achieving cell-type specific transgene expression typically rely on transgenic mouse lines. Cell-specific transgene expression cannot therefore be easily achieved in other model organisms, such as rats or monkeys, which may offer experimental advantages. Moreover, even when the mouse is a suitable model system, the generation and maintenance of new transgenic mouse lines is expensive, labor-intensive and slow. A general method for achieving specific cell-type expression of transgenes would therefore have many potential applications for research and therapy. We propose to develop a novel strategy for the expression of reporter genes in specific neuronal subtypes. Our strategy is based on the specific expression of either Cre or Flp recombinase in specific neuronal cell-types. The expression of transgenes can then be restricted to neurons that express the recombinase. However, unlike previous strategies in which cell-type specific recombinase expression is achieved through the generation of transgenic knock-in mice, our method can deliver both the recombinase and the transgene via recombinant viruses. Thus our method can be applied in organisms in which the potential for genetic manipulation is limited. Our approach will contribute to the understanding of the brain circuitry underlying addiction, and may eventually allow targeting of neuronal sub circuits in the treatment of human neuropsychiatric disorders.

描述（由申请人提供）：该项目的目的是开发一种广泛适用的方法，用于在大鼠，灵长类动物和其他哺乳动物的大脑中，在大鼠，灵长类动物和其他哺乳动物的大脑中进行方便的细胞类型特异性表达，例如光遗传学试剂。大脑中的电路由多种神经元亚型组成，这些神经元亚型通常由特定基因的选择性表达来定义。例如，由D1R和D2R多巴胺受体的表达定义的神经元代表不同的神经元亚群，这些神经元亚群在增强学习和成瘾中起着不同的作用。靶向越来越多的分子工具（包括粉醇和光遗传试剂）表达这种遗传学定义的神经元亚型的能力为解剖神经回路提供了一种有力的方法。但是，当前实现细胞类型特异性转基因表达的方法通常依赖于转基因小鼠系。因此，在其他模型生物（例如大鼠或猴子）中，无法轻松实现细胞特异性的转基因表达，这可能具有实验优势。此外，即使鼠标是合适的模型系统，新的转基因鼠标线的产生和维护也很昂贵，劳动力密集且缓慢。因此，一种实现转基因特异性细胞类型表达的一般方法将具有许多潜在的研究和治疗应用。我们建议制定一种在特定神经元亚型中表达报告基因的新策略。我们的策略基于特定神经元细胞类型中CRE或FLP重组酶的特定表达。然后可以将转基因的表达局限于表达重组酶的神经元。然而，与以前的策略不同，通过生成转基因敲门小鼠来实现细胞型特异性重组酶表达，我们的方法可以通过重组病毒既可以传递重组酶和转基因。因此，我们的方法可以应用于遗传操纵潜力受到限制的生物体中。我们的方法将有助于理解脑电路的基础成瘾，并最终允许靶向神经元亚电路治疗人类神经精神疾病。