Cloning and characterization of the Van Der Woude gene

Van Der Woude 基因的克隆和表征

• 批准号: 6929461
• 负责人: BRIAN C SCHUTTE
• 金额: $ 35.03万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-08-01 至 2010-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6929461
• 关键词: animal genetic material tag; biotechnology; cleft lip; cleft palate; clinical research; congenital oral /facial /cranial defect; developmental genetics; disease /disorder etiology; family genetics; gene environment interaction; gene expression; gene mutation; genetic mapping; genetic screening; genetically modified animals; high throughput technology; human genetic material tag; human subject; interferons; laboratory mouse; molecular cloning; polymerase chain reaction; single strand conformation polymorphism; syndrome; western blottings

DESCRIPTION (provided by applicant): Cleft lip and palate is a common congenital anomaly with significant lifelong morbidity. The extensive psychological, surgical, speech and dental involvement emphasize the importance of understanding the underlying causes. Our long-term objective is to discover the genes and environmental factors that contribute to this disorder. However, the etiology of cleft lip and palate is complex, and given the large number of genes predicted to be involved in isolated cleft lip and palate, gene discovery poses a significant challenge. The approach used here will be to study a genetically simple model of orofacial clefting, Van der Woude syndrome. Van der Woude syndrome is an outstanding clinical model for isolated cleft lip and palate, as its only distinguishing features are pits in the lower lip. Mutations in Interferon Regulatory Factor 6 (IRF6) cause Van der Woude syndrome. Moreover, genetic variation in IRF6 confers risk for isolated cleft lip and palate, demonstrating that Van der Woude syndrome is also an outstanding genetic model. Thus, IRF6 is essential for a critical pathway in palate development, and its discovery represents a foothold in this pathway. The main goal of this proposal is to delineate the IRF6 pathway by first determining the in vivo function of IRF6 and then identifying factors that regulate the expression of IRF6. There are 3 Aims: 1) To generate a mouse model for Van der Woude syndrome by mutating Irf6, and to perform a comprehensive analysis of the pathogenic processes in palate development in these mice. To investigate a role for Type I interferons in palate development in mice, as IRF6 is a member of the Interferon Regulatory family of transcription factors. 2) To evaluate a potential genetic interaction between mutations in Irf6 and Transformation growth factor beta3 (Tgfbeta3), since Tgfa3 is necessary for palate development and for Irf6 expression in the palate. Mice that are double heterozygotes for Irf6 and Tgfbeta3 will be analyzed for palate development. Also, to examine the potential role for Irf6 in transducing the Tgfbeta3 signal during palate development. 3) To identify the enhancer that regulates Irf6 expression in the palate using transgenic mice. Does Tgfbeta3 regulate Irf6 through the SMADs or by other transcription factors or both? Completion of these aims will advance our understanding for the role of IRF6 in palate development, and identify novel gene interactions and genes that may contribute to isolated cleft lip and palate, a common human disorder. This knowledge will provide the basis for enhanced diagnostic tests and future preventive interventions.

描述（由申请人提供）：唇腭裂是一种常见的先天性异常，具有显著的终身发病率。广泛的心理，手术，语言和牙科参与强调了解根本原因的重要性。我们的长期目标是发现导致这种疾病的基因和环境因素。然而，唇腭裂的病因学是复杂的，并且考虑到大量的基因预测参与孤立的唇腭裂，基因发现构成了重大挑战。这里使用的方法将是研究一个遗传学上简单的口面裂模型，货车德沃德综合征。货车der Woude综合征是孤立性唇腭裂的一个杰出的临床模型，因为其唯一的区别特征是下唇的凹陷。干扰素调节因子6（IRF 6）的突变导致货车德沃德综合征。此外，IRF 6的遗传变异赋予了孤立性唇腭裂的风险，这表明货车德沃德综合征也是一个杰出的遗传模型。因此，IRF6对于腭发育的关键途径是必不可少的，它的发现代表了这一途径的立足点。该提案的主要目标是通过首先确定IRF6的体内功能，然后确定调节IRF6表达的因子来描绘IRF6通路。有三个目标：1）通过突变Irf 6建立货车德沃德综合征小鼠模型，并对这些小鼠腭发育的致病过程进行全面分析。研究I型干扰素在小鼠腭发育中的作用，因为IRF6是转录因子干扰素调节家族的成员。2)评价Irf6和转化生长因子β 3（Tgf β 3）突变之间的潜在遗传相互作用，因为Tgfa3是腭发育和Irf6在腭中表达所必需的。将分析Irf 6和Tgf β 3双杂合子小鼠的腭发育。此外，研究在腭发育过程中Irf6在转导Tgf β 3信号中的潜在作用。3)使用转基因小鼠鉴定调节上颚中Irf6表达的增强子。Tgf β 3通过SMADs或其他转录因子或两者来调节Irf6吗？这些目标的完成将促进我们对IRF6在腭发育中的作用的理解，并确定新的基因相互作用和基因，可能有助于孤立的唇腭裂，一种常见的人类疾病。这些知识将为加强诊断测试和未来的预防性干预措施提供基础。