A new strategy for cell-type specific gene disruption in flies and mice

果蝇和小鼠细胞类型特异性基因破坏的新策略

基本信息

批准号：
9297370
负责人：
Thomas Robert Clandinin
金额：
$ 73.51万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-09-18 至 2019-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9297370
关键词：
Address Alleles Alpha Cell Animal Behavior Animal Model Behavior Behavioral Behavioral Assay Benchmarking Biological Assay Biological Models Brain Calcium Cells Clustered Regularly Interspaced Short Palindromic Repeats Code Collection Communities Complex DNA Insertion Elements DNA Transposable Elements DNA cassette Data Development Disease Drosophila genus Electrical Synapse Electrophysiology (science)Elements Enterobacteria phage P1 Cre recombinase Enzymes Future Gated Ion Channel Gene Expression Gene Expression Profile Gene Mutation Gene Targeting Genes Genetic Genetic Recombination Genetic Transcription Genome Genomics Goals Health Histologic Image Individual Instinct Introns Ion Channel Gating Label Lead Link Mammals Measures Mediating Membrane Potentials Molecular Mus Mutation Nervous system structure Neurodegenerative Disorders Neurons Neurosciences Neurotransmitter Receptor Neurotransmitters Output Pattern Pharmacology Physiological Play Process Proteins Reagent Reporter Reporting Research Personnel Role SNAP receptor Shapes Specificity Synapses Synaptic Transmission TPH2 Temperature Transgenic Organisms Tyrosine 3-Monooxygenase Validation Visual Visual system structure Work behavioral study brain dysfunction cell type design efficacy testing experience flexibility fly gene function in vivo interest member nervous system disorder neural circuit neural patterning neuronal circuitry neuronal excitability neurophysiology neuropsychiatric disorder neuroregulation neurotransmission public health relevance recombinase relating to nervous system tool treatment strategy two-photon vesicular GABA transporter vesicular glutamate transporter 2 virtual voltage

项目摘要

DESCRIPTION (provided by applicant): Developing and validating a toolkit for cell type specific gene manipulation PIs: Clandinin and Shah Neurons express complex arrays of genes that play crucial roles in determining neuronal function. As such, single-gene mutations can lead to neurodevelopmental, neurophysiological, and neurodegenerative diseases. However, the nervous system is made up of many different types of neurons, which differ both in the genes they express and the function those genes perform. The ability to inactivate targeted genes only in the cell type of interest is therefore critical for our understanding of neural circuit function This application will generate a generalizable, validated set of transgenic flies and mice for cell type specifically manipulating genes that control the inputs, outputs, and activity patterns of neurons in physiological and behavioral studies of a wide array of circuits. This application describes three goals for developing and validating this toolkit. First, we will generate conditional allelesof 24 genes important for neuronal excitability and signaling in Drosophila. Second, we will validate our cell type specific gene disruption in vivo using a combination of voltage imaging, calcium imaging and behavioral assays, providing strong evidence of the broad utility of this toolkit. Third, since mice are critical model organisms for studying vertebrate nervous systems in health and in disease, we will adapt the same tool we use in flies to mice, targeting a key set of genes controlling excitation, inhibition and neuromodulation. Development of this toolkit will provide th neuroscience community with the means to manipulate essential neural genes with unprecedented precision in cell types of interest, thereby allowing fundamental questions about how genes shape neuronal circuit function to be addressed. As mutations in these classes of genes lead to devastating neurological disorders, this tool will facilitate studies that expand our understanding of these diseases and the treatment possibilities. As pharmacological approaches to treating brain dysfunction are ultimately limited by molecular specificity, understanding cell-type specific gene function is critical to the development of new treatment strategies. Finally, as the tool we will develop can be generalized to virtually any gene, future studies can extend the use of this tool to any gene of interest, in either flies or mice.

描述（应用程序提供）：开发和验证细胞类型特定基因操纵的工具包PI：Clandinin和Shah神经元表达基因的复杂阵列，这些基因在确定神经元功能中起着至关重要的作用。因此，单基因突变会导致神经发育，神经生理学和神经退行性疾病。但是，神经系统由许多不同类型的神经元组成，它们在它们表达的基因和这些基因的功能中都不同。因此，仅在感兴趣的细胞类型中灭活靶向基因的能力对于我们对神经回路功能的理解至关重要专门操纵控制各种电路的物理和行为研究中神经元的输入，输出和活性模式的基因。该应用程序描述了开发和验证该工具包的三个目标。首先，我们将产生24个基因的条件等位基因对果蝇中的神经元刺激和信号传导很重要。其次，我们将使用电压成像，钙成像和行为评估的组合在体内验证细胞类型的特定基因破坏，从而为该工具包的广泛实用性提供了有力的证据。第三，由于小鼠是研究健康和疾病中脊椎动物神经系统的关键模型组织，因此我们将适应苍蝇中使用的相同工具，以针对控制兴奋，抑制和神经调节的一组关键基因。该工具包的开发将为神经科学界提供手段，以在感兴趣的细胞类型中以前所未有的精度操纵基本神经元基因，从而允许有关基因如何塑造神经元电路功能的基本问题。由于这些类别的基因突变导致毁灭性神经系统疾病，该工具将促进扩大我们的研究了解这些疾病和治疗可能性。由于药物治疗脑功能障碍最终受到分子特异性的限制，了解细胞类型特异性基因功能对于新治疗策略的发展至关重要。最后，由于我们将开发的工具可以推广到几乎任何基因，因此未来的研究可以将该工具的使用扩展到果蝇或小鼠中任何感兴趣的基因。