CLONING AND CHARACTERIZATION OF THE VAN DER WOUDE GENE

唇腭裂(CL/P)是一种主要的先天性结构异常，具有终生发病率和复杂的病因。广泛的心理、外科、言语和牙科介入强调了了解潜在原因的重要性。对于研究人员来说，唇腭裂和其他复杂的疾病一样，在确定导致其表型的多种遗传、环境和随机因素方面存在挑战。在这项提案中，我们将通过对一种遗传上简单的唇裂形式--范德伍德综合征(VDWS)的研究，来探讨唇腭裂的复杂原因。VDWS是最常见的唇腭裂综合征形式，我们认为最适合于增进对更常见的非综合征形式的唇腭裂的了解，我们认为最适合的形式有助于增加对更常见的非综合征形式的了解。该项目的具体目标将包括：1)VDWS基因的鉴定。致病微缺失的基因筛查将被用来进一步限制包含VDWS基因的区域。强大的基因发现技术，包括整个关键区域的序列分析，将被用来识别转录单位。然后，突变筛查将被用来发现引起DNA变化的疾病。将进行广泛的突变筛选，以寻找功能结构域。2)VDWS基因及其小鼠同源基因的特征，包括完整的cDNA和基因组序列分析，时间和组织特异性表达的研究，以确定需要VDWS基因功能的发育途径，并筛选可能在相同途径中发挥作用的基因同源基因。3)识别调控VDWS基因表达的序列和转基因小鼠模型的发展，包括将用于4)长期研究的小鼠敲除，这些研究将包括互补实验，以识别该途径中的其他基因，并调查环境因素对VDWS基因和同一途径中的其他基因的影响。这个项目的影响是，CL/P的研究是有价值的，既是因为缺陷本身作为发病率的贡献者的重要性，也是因为它为复杂的人类出生缺陷提供了一个模型。VDWS和VDWS样基因的识别将立即提供更好的风险咨询，它们在环境应激下的特征有望为预防策略和改进治疗做出贡献。此外，VDWS基因的鉴定与CL/P的研究相关，因为它与更常见的非综合征形式的CL/P相似，而且它将为颅面发育的早期阶段提供立足点。

Cleft lip and palate (CL/P) is a major congenital structural anomaly that is notable for significant lifelong morbidity and complex etiology.. The extensive psychological, surgical, speech and dental involvement emphasize the importance of understanding the underlying causes. For the investigator, cleft lip and palate, like other complex diseases, provides a challenge in determining the multiple genetic, environmental and stochastic factors that lead to its phenotype. In this proposal, we will pursue the complex causes of cleft lip and palate through our investigations of a genetically simple form of clefting, Van der Woude syndrome (VDWS). VDWS is the most frequent syndromic form of cleft lip and palate and the form we fell is best suited to contribute to an increased understanding of the more common non-syndromic form of cleft lip and palate and the form we feel is best suited to contribute to an increased understanding of the more common non-syndromic form. Specific goals in this project will include: 1) identification of the VDWS gene. A genetic screen for disease-causing micro-deletions will be used to further restrict the region that contains the VDWS gene. Powerful gene-finding techniques, including the sequence analysis of the entire critical region, will be used to identify transcriptional units. Mutation screens will then be used to find disease causing changes in the DNA. Extensive mutation screens will be performed to search for functional domains. 2) characterization of the VDWS gene and its mouse homolog, including complete cDNA and genomic sequence analysis, the study of temporal and tissue-specific expression to identify developmental pathways that require the VDWS gene functional and screens for gene homologs which may function in the same pathway. 3) identification of sequences that regulate VDWS gene expression and the development of transgenic mouse models, including a mouse knockout that will be used in 4) long-term studies that will include complementation experiments to identify other genes in the pathway and investigate the effects of environmental factors on the VDWS gene and other genes in the same pathway. The impact of this project is that studies of CL/P are valuable, both for the importance of the defect itself as a contributor to morbidity and for providing a model for a complex human birth defect. The identification of the VDWS and VDWS-like genes will immediately provide for better risk counseling, and their characterization under environmental stresses hold the promise for contribution to preventative strategies and improved therapeutics. In addition, the identification of the VDWS gene is relevant to studies of CL/P because of its similarities to the more frequent non-syndromic forms of CL/P and because it will provide a foothold into the earliest steps of craniofacial development.