Generation of mice with inducible Cre expression in adult cochlear hair cells

成年耳蜗毛细胞中具有诱导型 Cre 表达的小鼠的产生

基本信息

批准号：
7240216
负责人：
JIAN ZUO
金额：
$ 18.75万
依托单位：
ST. JUDE CHILDREN'S RESEARCH HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-02-10 至 2009-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7240216
关键词：
3&apos Untranslated Regions Adopted Adult Alleles Animal Model Animals Auditory Bacteriophages Cell Nucleus Cells Chimeric Proteins Deposition Development Disease Down-Regulation Ear Engineering Enhancers Ensure Estrogen Receptors Gene Expression Generations Genes Genetic Genome Hair Hair Cells Hearing Hearing Impaired Persons Hearing problem Human Individual Infant Inner Hair Cells Intervention Knockout Mice Laboratories Labyrinth LacZ Genes Life Live Birth Mus Mutant Strains Mice Mutate Myosin ATPase Noise-Induced Hearing Loss Outer Hair Cells Pathology Persons Physiology Play Process Public Health Regulatory Element Reporter Research Personnel Resources Retinoblastoma Genes Role Specificity Tamoxifen Time Transgenes Transgenic Mice Transgenic Organisms United States Untranslated Regions Variant Yeasts aged cell type day deafness hearing impairment interest mature animal mouse genome neurotrophic factor postnatal rat Pres protein recombinase spiral ganglion therapeutic target tool

项目摘要

DESCRIPTION (provided by applicant): Our ability to manipulate gene expression specifically in the hair cells of the inner ear during development and adulthood in mice is crucial for understanding the physiology of hearing and the pathology of deafness in humans. Recent advances from our laboratory and many others have demonstrated that gene expression can be manipulated in developing mouse hair cells in a spatially and temporally controlled manner. However, a key feature remains elusive -- our ability to manipulate gene expression specifically in mature inner hair cells (IHCs) and outer hair cells (OHCs). Here we propose to develop transgenic or knockin mice that express inducible CreERT2, an effective fusion of Cre and estrogen receptor (CreER), in mature cochlear hair cells (OHCs in Aim 1; IHCs and OHCs in Aim 2). The characterization and availability of these mouse lines will enable auditory researchers to inactivate or activate genes of their interest in mature hair cells. The creation of conditional (loxP) alleles of every mouse gene in their genome, which has been proposed in the NIH's Knockout Mouse Project (KOMP), will provide a complementary resource for the use of the inducible Cre lines that we generate. Finally, the strategies we adopt here can be used to express genes specifically in other mature cell types of the inner ear (i.e., IHCs, spiral ganglia, stria fibrocytes and marginal cells) at any given time. All mouse lines will be genetically engineered in 129S7/C57BL/6J mixed or FVB/NJ strains, transferred to the CBA/CaJ strain, and then deposited in the NIH-sponsored Mutant Mouse Regional Resource Center (MMRRC). Public Health Relevance Statement An estimated 28 million people in the United States are deaf or hard of hearing. Approximately 1.5 million individuals aged 3 years or older are deaf in both ears and 2 to 3 per 1,000 live births suffer congenital hearing loss. More than 40 million persons in the United States suffer various levels of noise induced hearing loss. Despite the significant progress in our understanding of these hearing disorders, very little is known about the disease causes and about the normal hearing processes in adults. Many genetic factors (genes) that, when mutated, cause hearing impairment in people, play critical roles in both infants and adults. We propose here to develop genetic tools that would allow us to investigate the function of hearing related genes in adult animal models. These tools will be able to surgically activate genes in a specific set of cells in the hearing process at any time of the adult life of the animal. This temporal and spatial precision is critical for our understanding of hearing process and developing therapeutic targets for intervention of the hearing disorders.

描述（由申请人提供）：我们在小鼠发育和成年期间特异性操纵内耳毛细胞中的基因表达的能力对于理解人类听力生理学和耳聋病理学至关重要。我们实验室和许多其他实验室的最新进展表明，可以以空间和时间控制的方式操纵小鼠毛细胞发育中的基因表达。然而，一个关键特征仍然难以捉摸——我们特异地操纵成熟内毛细胞 (IHC) 和外毛细胞 (OHC) 中基因表达的能力。在这里，我们建议开发在成熟耳蜗毛细胞（目标 1 中的 OHC；目标 2 中的 IHC 和 OHC）中表达可诱导的 CreERT2 的转基因或敲入小鼠，CreERT2 是 Cre 和雌激素受体 (CreER) 的有效融合。这些小鼠品系的表征和可用性将使听觉研究人员能够灭活或激活成熟毛细胞中他们感兴趣的基因。 NIH 的基因敲除小鼠计划 (KOMP) 中提出，为小鼠基因组中的每个基因创建条件 (loxP) 等位基因，将为我们生成的诱导型 Cre 系的使用提供补充资源。最后，我们在此采用的策略可用于在任何给定时间在内耳的其他成熟细胞类型（即 IHC、螺旋神经节、纹状纤维细胞和边缘细胞）中特异性表达基因。所有小鼠品系都将在 129S7/C57BL/6J 混合品系或 FVB/NJ 品系中进行基因工程改造，转移至 CBA/CaJ 品系，然后存放在 NIH 资助的突变小鼠区域资源中心 (MMRRC)。公共卫生相关性声明据估计，美国有 2800 万人耳聋或有听力障碍。大约 150 万 3 岁或以上的人双耳失聪，每 1,000 名活产儿中就有 2 至 3 人患有先天性听力损失。美国有超过 4000 万人患有不同程度的噪音引起的听力损失。尽管我们对这些听力障碍的理解取得了重大进展，但对疾病的原因和成人的正常听力过程知之甚少。许多遗传因素（基因）在突变时会导致人类听力障碍，对婴儿和成人都起着至关重要的作用。我们在此建议开发遗传工具，使我们能够研究成年动物模型中听力相关基因的功能。这些工具将能够在动物成年后的任何时间通过手术激活听力过程中一组特定细胞中的基因。这种时间和空间精度对于我们理解听力过程和制定听力障碍干预治疗目标至关重要。