Genetic Targeting Core

基因靶向核心

基本信息

批准号：
10626083
负责人：
Edita Navratilova
金额：
$ 37.3万
依托单位：
UNIVERSITY OF ARIZONA
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-08-15 至 2026-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10626083
关键词：
Amygdaloid structure Anatomy Animal Experiments Applications Grants Area Basal Ganglia Behavioral Brain Brain region CRISPR/Cas technology Cannabinoids Capsid Cells Central Nervous System Chronic Collaborations Complex Corticotropin-Releasing Hormone Cre driver Data Development Dopamine Drug usage Drug user Dynorphins Endocannabinoids Engineering Ensure Experimental Designs Future G-Protein-Coupled Receptors Gene Deletion Gene Delivery Gene Mutation Gene Targeting Genes Genetic Genetic Materials Genetic Techniques Genome Glutamates Goals Habits Halorhodopsins Immunohistochemistry Impairment Individual Investigation Ion Channel Knowledge Light Methodology Methods Microscopy Modernization Mus National Institute of Drug Abuse Neurons Neurotransmitters Non-Viral Vector Norepinephrine Opioid Opioid Peptide Opsin Outcome Outcome Measure Peripheral Nervous System Pharmaceutical Preparations Phase Plasmid Cloning Vector Plasmids Prefrontal Cortex Prevention strategy Property Proteins Protocols documentation Rattus Relapse Reporter Genes Reproducibility Research Research Personnel Resources Rewards Rodent Role Serotonin Services Signal Pathway Specificity Standardization Stress Substance Use Disorder System Techniques Technology Transduction Gene Transfection Transgenes Transgenic Mice Transgenic Organisms Viral Viral Vector Visualization Withdrawal Work addiction brain circuitry cell type design designer receptors exclusively activated by designer drugs executive function genetic approach improved in vivo inducible gene expression innovation instrument microbial neural neural circuit neuromechanism neurotransmission novel optogenetics overexpression pharmacologic prevent restoration substance use treatment tool transgene expression

项目摘要

Project Summary – Genetic Targeting Core Substance use disorders (SUD) are characterized by dysregulation of brain circuitry that involves diminished activity of the brain reward circuits, increased responsiveness of the stress circuits and impaired functioning of the executive cortical circuits. Neural changes are observed in the basal ganglia, extended amygdala and prefrontal cortical regions and encompass a wide range of endogenous neurotransmitters including dopamine, opioid peptides, endocannabinoids, corticotropin releasing factor (CRF), dynorphin, glutamate and others. It implies that these maladaptations may be causally responsible for behavioral changes often observed in chronic drug users, for example habit formation and compulsive drug taking, that further worsen the neuronal and behavioral signs of SUD. Restoration of normal functioning of the brain circuits is therefore a desirable goal of SUD therapies. However, the exact influence of chronic drug use on the addiction brain circuits and how manipulation of these circuits could prevent or treat substance use disorders remains to be elucidated. As part of the Center of Excellence in Addiction Studies, the Genetic Targeting Core (GT Core) will assist users in obtaining preliminary data on neural mechanisms of addiction that will form basis for future NIDA grant applications. The GT Core will provide services to target and manipulate the addiction circuits in a cell specific and circuits specific manner using state-of-the-art genetic techniques, including optogenetics, chemogenetics and CRISPR/Cas9 gene editing. Optogenetic strategies use microbial-based light activated ion channels (opsins) that allow fast neuronal activation (channelrhodopsins) or inhibition (halorhodopsins, archaerhodopsins), while chemogenetic approaches use engineered ion channels (PSAMs) or G protein- coupled receptors (DREADDs) that can be activated pharmacologicaly. CRISPR/Cas9 gene editing allows direct in vivo manipulation of the genome in rodents. The introduction of functional transgenes, such as opsins and DREADDs or editing of native genes requires in vivo delivery and expression of genetic material in desired cells. This is accomplished with viral or nonviral vector systems. Cell and circuit specificity can be further enhanced by the use of a specific Cre-driver mouse line. The GT Core will provide three main services to its users that correspond with the three Specific Aims: 1) assist with the selection of gene targeting methods, including the choice of the transgenic mouse line, and the type of viral vector; 2) optimize and implement gene delivery using high precision stereotaxic instruments; and 3) verify the efficacy of gene targeting using fluorescent microscopy or protein and gene quantification techniques. The GT Core will coordinate work with the Administrative, Behavioral and Neuroanalytical Cores to develop effective experimental strategies that involve several Cores. The close collaboration between our Cores ensures high expertise in all areas of the addiction research and will permit establishment of standardized outcome measures emphasizing scientific rigor and reproducibility.

项目摘要 - 遗传靶向核心药物使用障碍（SUD）的特征是脑电路的失调，涉及减少大脑的活动奖励电路，增加压力电路的响应能力以及功能受损执行皮层圈。在基底神经节，杏仁核和前额叶皮质区域，包括多巴胺，包括多巴胺，包括多巴胺的各种内源性神经递质阿片肽，内源性大麻素，皮质激素释放因子（CRF），dynorphin，谷氨酸等。它暗示这些不适可能是对经常在慢性中观察到的行为变化的因果关系吸毒者，例如习惯形成和强迫性药物，这进一步更糟的是神经元和 SUD的行为迹象。因此，恢复脑电路正常功能是一个理想的目标 SUD疗法。但是，慢性药物使用对成瘾脑回路的确切影响以及如何对这些电路的操纵可以预防或治疗药物使用障碍尚待阐明。作为成瘾研究卓越中心的一部分，遗传靶向核心（GT Core）将帮助用户在获取有关成瘾神经机制的初步数据时，将构成未来NIDA赠款的基础申请。 GT核心将为目标和操纵特定的单元格中的成瘾电路提供服务以及使用最先进的遗传技术（包括光遗传学，化学遗传学）的特定方式和CRISPR/CAS9基因编辑。光遗传学策略使用基于微生物的光激活离子通道（Opsins）允许快速神经元激活（通道Rhopopsins）或抑制作用（Halorhopopsins，，，， Archaerhopopsin），而化学遗传学方法使用工程离子通道（PSAM）或G蛋白可以通过药物激活的耦合受体（可怕）。 CRISPR/CAS9基因编辑允许直接体内对啮齿动物中基因组的操纵。引入功能翻译，例如Opsins和可怕的或天然基因的编辑需要体内递送和所需细胞中遗传物质的表达。这是通过病毒或非病毒载体系统实现的。单元和电路特异性可以通过使用特定的CRE驱动器鼠标线。 GT核心将为其用户提供三个主要服务，与三个特定目的相对应：1）协助选择基因靶向方法，包括选择转基因小鼠系的选择，以及病毒载体； 2）使用高精度的立体定位仪器优化和实施基因传递； 3）验证使用荧光显微镜或蛋白质和基因定量技术靶向基因的效率。 GT核心将与行政，行为和神经分析核心协调工作，以发展有效涉及多个核心的实验策略。我们的核心之间的密切合作确保了高度成瘾研究的所有领域的专业知识，并将允许建立标准化的结果指标强调科学严格和可重复性。