Characterization of NSCLP gene CRISPLD2

描述（由申请人提供）：非综合征性唇裂伴或不伴腭裂（NSCLP）是一种常见的复杂出生障碍，影响1/700的活产婴儿，在美国每年有4000例。NSCLP的病因尚不完全清楚，但已知遗传和环境影响都涉及其中。估计有12-25%的遗传变异导致NSCLP已被确定。我们最近发现，一种新的基因CRISPLD2与NSCLP有关，并在发育中的小鼠胚胎的口面部区域表达，此时的唇腭裂形成。我们假设CRISPLD2在颅面发育过程中起重要作用，该基因的变异可能导致非小细胞肺癌。人们对这种新基因知之甚少。该项目的目标是表征该基因在正常颅面发育中的作用，以及它如何促进NSCLP。为了实现这一目标，将对CRISPLD2序列进行变异研究。确定的序列变异将使用计算机算法评估预测功能，然后在体外分析以确定对正常基因功能的影响。这种模式已被证明在识别其他复杂人类疾病（如巨结肠病）和其他NSCLP基因（如Msx1）的致病变异方面是成功的，并将被用于识别具有潜在生物学功能的CRISPLD2序列变异。为了确定CRISPLD2在正常脊椎动物发育中的功能，将使用条件构建来敲除小鼠中的CRISPLD2。小鼠模型已被证明可用于研究其他裂裂基因（如WntQb、IRF6和Msx1）的基因功能。这些研究是表征这种新的颅面基因的重要的第一步。本研究将有助于我们更好地了解颅面发育和NSCLP的病因。该项目达到了国家口腔疾病防治中心通过研究、研究培训和传播健康信息改善口腔健康的目标。非小细胞肺癌患者在外科、牙科和语言治疗方面面临着巨大的医疗负担。从该项目中获得的信息将确定我们最近发现的新裂解基因CRISPLD2的功能，并定义可能导致NSCLP的基因变化。这将有助于理解正常的面部发育，有助于更好地诊断和咨询有唇裂风险的家庭，并可能导致对唇裂的治疗干预。

DESCRIPTION (provided by applicant): Nonsyndromic cleft lip with or without cleft palate (NSCLP) is a common complex birth disorder that affects 1/700 live births, with 4000 cases each year in the United States. The etiology of NSCLP is not completely understood, but both genetics and environmental influences are known to be involved. An estimated 12-25% of the genetic variation causing NSCLP has been identified. We have recently shown that a novel gene, CRISPLD2, is associated with NSCLP and expressed in the orofacial region of developing mouse embryos at the time of lip and palate formation. We hypothesize that CRISPLD2 plays an important role during craniofacial development and that variation in this gene may cause NSCLP. Little information is known about this novel gene. The goal of this project is to characterize the role of this gene in normal craniofacial development and and how it contributes to NSCLP. To accomplish this goal, the CRISPLD2 sequence will be interrogated for variation. Identified sequence variants will be assessed for predicted function using computer algorithms and then analyzed in vitro to determine the effect on normal gene function. This paradigm has proven successful in identifying causative variants in other complex human diseases (i.e., Hirschsprung disease) and other NSCLP genes (i.e., Msx1), and will be followed to identify CRISPLD2 sequence variants with a potential biologic function. In order to determine CRISPLD2 function in normal vertebrate development, a conditional construct will be used to knock out CRISPLD2 in a mouse. Mouse models have proven useful in studying gene functions of other clefting genes (i.e., WntQb, IRF6 and Msx1). These studies are important first steps to characterize this novel craniofacial gene. This research will help us gain a better understanding craniofacial development and the causes of NSCLP. This project meets the goals of the NIDCR in improving oral health through research, research training and the dissemination of health information. Patients with NSCLP face a significant healthcare burden from surgical, dental and speech therapies. The information gained from this project will identify the function of the new clefting gene, CRISPLD2, that we recently identified and define the changes in the gene that may contribute to NSCLP. This will aid in the understanding of normal facial development, help to better diagnose and counsel families at risk for having a child with a cleft, and may lead to therapeutic intervention of orofacial clefting.