Transgenic mice containing human DUF1220 domains

含有人 DUF1220 结构域的转基因小鼠

基本信息

批准号：
8130843
负责人：
JAMES M SIKELA
金额：
$ 14.49万
依托单位：
UNIVERSITY OF COLORADO DENVER
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-08-18 至 2012-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8130843
关键词：
Affect Animal Model Antibodies Autistic Disorder Behavior Behavioral Brain Breeding Clip Code Cognition Disorders Cognitive Copy Number Polymorphism DNA Disease Engineering Evolution Foundations Gene Expression Generations Genes Genetic Markers Genetic Transcription Goals Human Human Genome Human Identifications Injection of therapeutic agent Learning Mammals Memory Mental Retardation Messenger RNA Microcephaly Modeling Molecular Molecular Profiling Monitor Mus Neocortex Neurons Pattern Phase Phenotype Presynaptic Terminals Primates Process Production Proteins Reporting Research Project Grants Resolution Role Schizophrenia Series Site Structure Tail Tertiary Protein Structure Testing Tissues Tomatoes Transgenes Transgenic Mice Transgenic Organisms Western Blotting Work awake behavior test cognitive function design dosage follow-up human tissue immunocytochemistry mouse genome mouse model neuronal cell body promoter public health relevance research study response tau Proteins tool

项目摘要

DESCRIPTION (provided by applicant): DUF1220 protein domains are highly amplified in the human lineage, show signs of positive selection and, in brain, are highly expressed in regions associated with higher cognitive function. Virtually all DUF1220 domains are primate-specific, with the highest copy number (212) found in humans; mice have only one copy and none of the primate type. The majority of DUF1220 domain sequences are found at 1q21.1, and several reports have found copy number variations in 1q21.1 associated with a number of cognitive diseases including autism, schizophrenia and microcephaly. Given the potential importance of DUF1220 to human cognitive disease and brain evolution, the goal of this application is to develop the first animal model encoding human DUF1220 domains. Two human BAC clones each encoding a different human DUF1220 gene and multiple DUF1220 domains, will be used for the production of transgenic mice. Additional sequences (tauGFP and tau/Tomato) will be added to each construct to allow expression of the gene to be followed in both neuronal cell bodies and axonal projections. The mice will be initially characterized by Western analysis using the anti- DUF1220 antibody we have developed that sees the primate/human-specific DUF1220 domains but not the single DUF1220 domain found in mice. For follow-up characterization, those mice expressing the two DUF1220 transgenes will be used 1) for cross-breeding experiments to increase the DUF1220 copy number to a more human- like level, 2) for high-resolution immunocytochemical analysis of brain expression patterns, and 3) for a series of behavioral tests designed to monitor learning and memory abilities. PUBLIC HEALTH RELEVANCE: This work should generate the first animal model to study the neuronal and behavioral function of DUF1220 domains, sequences that may be important to higher brain function in humans. As a result, these models should increase our understanding of the molecular mechanisms that underlie certain cognitive diseases such as mental retardation and autism.

描述（由申请人提供）：DUF1220蛋白结构域在人类谱系中高度扩增，显示出正选择的迹象，并且在大脑中，在与较高认知功能相关的区域中高度表达。几乎所有 DUF1220 结构域都是灵长类动物特异性的，其中在人类中发现的拷贝数最高 (212)；小鼠只有一份，而灵长类动物则没有。大多数 DUF1220 结构域序列位于 1q21.1，并且一些报告发现 1q21.1 中的拷贝数变异与许多认知疾病（包括自闭症、精神分裂症和小头畸形）相关。鉴于 DUF1220 对人类认知疾病和大脑进化的潜在重要性，本申请的目标是开发第一个编码人类 DUF1220 结构域的动物模型。两个人类 BAC 克隆各自编码不同的人类 DUF1220 基因和多个 DUF1220 结构域，将用于生产转基因小鼠。额外的序列（tauGFP 和 tau/Tomato）将被添加到每个构建体中，以允许在神经元细胞体和轴突投射中跟踪基因的表达。小鼠最初将使用我们开发的抗 DUF1220 抗体进行蛋白质印迹分析，该抗体可以看到灵长类/人类特异性的 DUF1220 结构域，但不能看到小鼠中发现的单个 DUF1220 结构域。对于后续表征，表达两种 DUF1220 转基因的小鼠将用于 1) 杂交实验，以将 DUF1220 拷贝数增加到更接近人类的水平，2) 用于大脑表达模式的高分辨率免疫细胞化学分析， 3) 一系列旨在监测学习和记忆能力的行为测试。公共健康相关性：这项工作应该产生第一个动物模型来研究 DUF1220 结构域的神经元和行为功能，这些序列可能对人类高级大脑功能很重要。因此，这些模型应该会增加我们对某些认知疾病（例如智力低下和自闭症）背后的分子机制的理解。