Transgenic mice containing human DUF1220 domains

描述(申请人提供)：DUF1220蛋白结构域在人类谱系中高度扩增，显示出积极选择的迹象，在大脑中，与较高认知功能相关的区域高度表达。几乎所有的DUF1220结构域都是灵长类特有的，在人类中发现的拷贝数最高(212)；老鼠只有一个拷贝，没有灵长类的类型。大多数DUF1220结构域序列位于1q21.1，一些报告发现1q21.1的拷贝数变化与许多认知疾病有关，包括自闭症、精神分裂症和小头畸形。鉴于DUF1220对人类认知疾病和脑进化的潜在重要性，这项应用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克隆将用于生产转基因小鼠，每个克隆编码不同的人类DUF1220基因和多个DUF1220结构域。额外的序列(tauGFP和tau/番茄)将被添加到每个构建中，以允许基因在神经元胞体和轴突投射中的表达。利用我们开发的抗DUF1220抗体对小鼠进行初步的Western分析，这种抗体可以看到灵长类/人类特有的DUF1220结构域，但不能看到在小鼠中发现的单个DUF1220结构域。为了后续的鉴定，这些表达两个DUF1220转基因基因的小鼠将被用于1)杂交实验，以将DUF1220拷贝数增加到更接近人类的水平，2)用于高分辨率免疫细胞化学分析大脑表达模式，以及3)用于一系列旨在监测学习和记忆能力的行为测试。 公共卫生相关性：这项工作应该建立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更高的大脑功能很重要。因此，这些模型应该会增加我们对某些认知疾病(如智力低下和自闭症)的分子机制的理解。

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.