Cloning and characterization of the Van Der Woude gene

Van Der Woude 基因的克隆和表征

• 批准号: 7101793
• 负责人: BRIAN C SCHUTTE
• 金额: $ 34.21万
• 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/7101793
• 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）：唇裂和腭裂是一种常见的先天性异常，具有显着的终生发病率。广泛的心理、外科、言语和牙科参与强调了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，唇裂和腭裂的病因很复杂，并且考虑到预测与孤立性唇裂和腭裂有关的大量基因，基因发现提出了重大挑战。这里使用的方法将是研究口面裂的遗传简单模型，即范德沃德综合征。范德沃德综合征是孤立性唇裂和腭裂的杰出临床模型，因为其唯一的显着特征是下唇凹陷。干扰素调节因子 6 (IRF6) 突变会导致范德沃德综合征。此外，IRF6 的遗传变异会带来孤立性唇裂和腭裂的风险，这表明范德沃德综合征也是一种出色的遗传模型。因此，IRF6 对于味觉发育的关键途径至关重要，它的发现代表了该途径的立足点。该提案的主要目标是通过首先确定 IRF6 的体内功能，然后识别调节 IRF6 表达的因子来描绘 IRF6 通路。目标有3个：1）通过Irf6突变建立Van der Woude综合征小鼠模型，并对这些小鼠上颚发育的致病过程进行全面分析。研究 I 型干扰素在小鼠上颚发育中的作用，因为 IRF6 是干扰素调节转录因子家族的成员。 2) 评估 Irf6 突变和转化生长因子 beta3 (Tgfbeta3) 之间潜在的遗传相互作用，因为 Tgfa3 对于上颚发育和 Irf6 在上颚中的表达是必需的。将分析 Irf6 和 Tgfbeta3 双杂合子小鼠的上颚发育。此外，还研究了 Irf6 在上颚发育过程中转导 Tgfbeta3 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。 Tgfbeta3 是否通过 SMAD 或其他转录因子或两者调节 Irf6？这些目标的完成将增进我们对 IRF6 在上颚发育中的作用的理解，并确定新的基因相互作用和可能导致孤立性唇裂和腭裂（一种常见的人类疾病）的基因。这些知识将为加强诊断测试和未来的预防干预措施提供基础。