Res Support Core

资源支持核心

基本信息

批准号：
10386843
负责人：
Guoping Feng
金额：
$ 75.32万
依托单位：
HARVARD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-09-05 至 2024-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10386843
关键词：
ASD patient Animal Model Behavior monitoring Behavioral Biological Sciences Biomedical Research Brain Brain Diseases CRISPR/Cas technology Calcium Callithrix Cell model Cells Collaborations Communities Complex Core Facility Defect Development Disease Disease model Electrophysiology (science)Equipment Evolution Eye Functional disorder Funding Genetic Genetic Engineering Goals Grant Human Image Impaired cognition Induced pluripotent stem cell derived neurons Infant Institutes Knock-in Knock-in Mouse Knock-out Knockout Mice Label Laboratories Massachusetts Mediating Mental disorders Modeling Monitor Mood Disorders Mus Mutation Neurodevelopmental Disorder Neurons Neurosciences Organoids Parents Pathologic Processes Physiology Prefrontal Cortex Primates Process Property Reaction Research Research Personnel Research Support Resources Reversal Learning Rodent Rodent Model Schizophrenia Social Interaction Staging Structure Study models System Testing Transgenic Mice Universities Viral Work behavior test brain tissue cell type cognitive function gaze gene function genetic manipulation human embryonic stem cell line human stem cells mouse genome mouse model mutant neurobiological mechanism neuron development neuropathology optogenetics preference repetitive behavior tool

项目摘要

Abstract The work to be done in the laboratory of Dr. Guoping Feng at MIT covers the following activities: 1. Provide Shank3 ASD mouse models and assistance for investigating PV neuron dysfunction. In addition to the Shank3 knockout mice we originally shared with Takao Hensch, we have recently developed two new Shank3 models: a conditional Shank3 knockin mouse that allows us to restore Shank3 expression at any developmental stages with Cre-ER, and a conditional Shank3 knockout mouse that allows us to delete Shank3 at any developmental stages with Cre-ER. We will provide these mice for a variety of structural and functional analysis. 2. Develop and provide genetic tools and resources for studying PV neuron function in marmosets. As a Core Facility, we will provide marmoset brain tissues of various developmental stages for studying PV neuron development and function in a primate brain. We will develop GABAergic neuron- specific AAV system for gene manipulations and calcium imaging. Most importantly, we will provide PV neuron-specific knockin marmoset (on-going effort) for viral-mediated, Cre-dependent manipulation of gene function and neuronal imaging in PV neurons. 3. Develop and characterize marmoset models for studying PV neuron dysfunction in ASD. A recent collaborative work between the laboratories of Hensch and Feng using mouse models suggested that PV neuron defects may be a common neuropathology in ASD. Here we propose to further this study by generating and characterizing Shank3 knockout marmosets. These models will allow us to study PV neuron dysfunction in a well-developed primate prefrontal cortex but also dissect cellular and circuitry basis of higher brain function that are disrupted in severe ASD such as eye-gazing, social interaction and cognitive impairment. 4. Provide human stem cells with Shank3 mutations for studying human PV neurons in organoids. In collaboration with Kevin Eggan’s group at Harvard and Lindy Barret at Stanley Center for Psychiatric Research at Broad Institute, we have developed human ES cell lines with Shank3 mutations using CRISPR/Cas9 system. We will provide these cells to Paola Arlotta’s group for developing organoids.

抽象的在麻省理工学院的Guoping Feng博士实验室要做的工作涵盖以下活动： 1。提供Shank3 ASD小鼠模型和研究PV神经元功能障碍的帮助。除了我们最初与塔科·汉斯（Takao Hensch）共享的shank3淘汰小鼠外，我们最近还开发了两种新的shank3型号：有条件的shank3敲蛋白鼠标，使我们能够还原shank3 Cre-er的任何发育阶段的表达，以及有条件的shank3敲除鼠标允许我们在CRE-ER的任何发展阶段删除Shank3。我们将为A提供这些老鼠各种结构和功能分析。 2。开发并提供用于研究摩尔果中光伏神经元功能的遗传工具和资源。作为核心设施，我们将提供各种发育阶段的摩尔莫塞脑脑组织研究原发性大脑中的PV神经元的发育和功能。我们将发展GABA能神经元 - 特定的AAV系统用于基因操作和钙成像。最重要的是，我们将提供PV 神经特异性的敲蛋白果仁糖（持续的努力），用于病毒介导的Cre依赖性基因操纵 PV神经元中的功能和神经元成像。 3。开发和表征用于研究ASD中PV神经元功能障碍的Marmoset模型。使用鼠标模型的汉施和冯实验室之间的最新合作工作建议PV神经元缺陷可能是ASD中常见的神经病理学。在这里我们建议进一步这项研究通过产生和表征shank3敲除果果。这些模型将使我们能够在发达的原发前额叶皮层中研究PV神经元功能障碍，但也剖析细胞和在严重的ASD中破坏大脑功能的电路基础，例如凝视，社交互动和认知障碍。 4。为人类干细胞提供shank3突变，用于研究器官中的人类PV神经元。与哈佛大学的凯文·埃格甘（Kevin Eggan）和斯坦利中心的林迪·巴雷特（Lindy Barret）合作 Broad Institute的精神病研究，我们开发了具有Shank3突变的人类ES细胞系使用CRISPR/CAS9系统。我们将向Paola Arlotta的小组提供这些细胞，用于开发类器官。