Characterization of NSCLP gene CRISPLD2

描述(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疾病，影响1/700活产，在美国每年有4000例。NSCLP的病因尚不完全清楚，但已知与遗传和环境影响有关。据估计，引起NSCLP的遗传变异中有12%-25%已被识别。我们最近发现了一个新的基因CRISPLD2，它与NSCLP相关，并在唇腭部形成时发育中的小鼠胚胎的口面部表达。我们推测CRISPLD2在颅面发育过程中起重要作用，该基因的变异可能导致NSCLP。人们对这种新基因知之甚少。该项目的目标是确定该基因在正常头面部发育中的作用，以及它在非小细胞肺癌中的作用。为了实现这一目标，将询问CRISPLD2序列是否存在变异。识别出的序列变异将使用计算机算法评估预测功能，然后在体外进行分析，以确定对正常基因功能的影响。这一模式已被证明在其他复杂人类疾病(如先天性巨结肠)和其他NSCLP基因(如Msx1)中成功识别致病变异，并将被用于识别具有潜在生物功能的CRISPLD2序列变异。为了确定CRISPLD2在脊椎动物正常发育中的功能，将使用一个条件结构来敲除小鼠的CRISPLD2。小鼠模型已被证明在研究其他分裂基因(即WntQb、IRF6和Msx1)的基因功能方面是有用的。这些研究是表征这种新的头面部基因的重要的第一步。这项研究将有助于我们更好地了解颅面发育和NSCLP的原因。该项目符合NIDCR通过研究、研究培训和传播健康信息来改善口腔健康的目标。NSCLP患者面临着外科、牙科和语言治疗带来的巨大医疗负担。从这个项目中获得的信息将确定我们最近发现的新的分裂基因CRISPLD2的功能，并确定可能与NSCLP有关的基因变化。这将有助于了解正常的面部发育，有助于更好地诊断和咨询患有唇裂儿童的风险家庭，并可能导致口腔唇裂的治疗干预。

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.