Mutagenesis and Murine Embyonic Development

描述（由申请人提供）：我们已经承担了一个项目，通过筛选ENU诱变小鼠的隐性突变来产生人类先天性缺陷模型， 胚胎发育后期该屏幕包含了一个基因图谱组件，目的是 以便于突变基因的定位克隆和功能表征。该策略效果良好，我们已经产生了许多表型类似的突变株系， 人类畸形综合征和出生缺陷。迄今为止发现的异常谱是显著不同的;例如，我们已经生成了脊椎肋骨发育不全、Robin序列、先天性骨缺损、非综合征性腭裂、多囊肾病、表皮大疱、非大疱性先天性鱼鳞病样红皮病和结构性心脏病的模型。我们已经绘制了其中的一些，并确定了9行的突变位点。为了实现这一目标，我们利用了有效的遗传作图和定位克隆技术。我们已经确定的许多基因的功能还没有得到很好的理解，我们已经发起了各种生化和发育研究来探索它们。除了表征突变小鼠中缺陷的生物学特性外，我们还在几个案例中确定了我们鉴定的基因在人类疾病的病因中起作用。因此，作为我们最初建议基础的所有前提都得到了实验验证。在这一延续性提案中，我们希望改进筛选的特异性和灵敏度，并优化分析的几个方面，同时保持迄今为止证明如此富有成效的基本方法。

DESCRIPTION (provided by applicant): We have undertaken a project to generate models of human congenital defects by screening ENU-mutagenized mice for recessive mutations affecting late embryonic development. The screen incorporated a genetic mapping component, with an aim to facilitate the positional cloning and functional characterization of the mutant genes. The strategy has worked well, and we have generated many mutant lines with phenotypes similar to human malformation syndromes and birth defects. The spectrum of abnormalities found to date is remarkably varied; for example, we have generated models of spondylocostal dysostosis, Robin sequence, congenital diaphragmatic defect, non-syndromic cleft palate, polycystic kidney disease, epidermal bullosa, non-bullosa congenital icthyosiform erythroderma, and structural heart disease. We have mapped a number of these, and identified the mutated locus in 9 lines. To accomplish this, we have taken advantage of efficent technologies for genetic mapping and positional cloning. The functions for many of the genes we have identified are not well understood, and we have initated a variety of biochemical and developmental studies to explore them. In addition to characterizing the biology of the defects in the mutant mice, we have in several cases established that the genes we identified play a role in the causation of human disease. Thus, all of the premises that were the basis of our original proposal have been experimentally validated. In this continuation proposal we hope to refine the specificity and sensitivity of the screen and optimize several aspects of the analysis, while maintaining the fundamental approach that has thus far proven so productive.