Development of on-chip ultra high-throughput whole-animal assay technologies

近年来，利用线虫线虫的优势 作为人类疾病的模型系统已经变得越来越明显，最终 在过去五年内两次获得诺贝尔生理学和医学奖。现有 脊椎动物模型，以及用于研究它们的仪器，不能 用于快速鉴定新基因和药物的高通量分析 线索。模式生物秀丽线虫允许在体内进行全基因组分析和高密度的 吞吐量筛选由于无与伦比的基因技术的可用性，其 透明度，以及以极小的数量增长的能力。然而，自从第一批出版物发表以来 在20世纪60年代初，科学家们手动操纵这种微小有机体的方式几乎没有改变， 因此，即使是简单的大规模化验也需要几个月到几年的时间 完成。重要的是，由于缺乏关键技术，几种检测方法也无法 对于高吞吐量的屏幕，要么完全执行，要么必须大大简化。 我们建议(1)开发第一个片上超高通量全流水线。 用于体内药物筛选和全基因组发现的动物操纵技术 基因功能和相互作用的复杂芯片行为和生化分析 策略；以及(2)首次大规模的神经机制活体研究。 使用建议的可复制损伤后的退化和再生 技巧。建议的技术和高通量分析策略可以 使用疾病模型显著影响许多分子机制的发现 小动物。

In recent years, the advantages of using the nematode Caenorhabditis elegans as a model system for human disease have become increasingly apparent, culminating in two Nobel Prizes in Physiology and Medicine within the last five years. Existing vertebrate animal models, and the instrumentation incorporated to study them, cannot be utilized for high throughput assays for rapid identification of novel genes, and drug leads. The model organism C. elegans allows in vivo genome-wide assays and high- throughput screens due to the availability of unmatched genetic techniques, its transparency, and the ability to grow it in minute volumes. Yet, since the first publications in early 1960s, little has changed how scientists manipulate this tiny organism manually, as a result of which even simple large-scale assays still take months to years to complete. Importantly, due to the lack of key technologies, several assays either cannot be performed at all or have to be dramatically simplified for high-throughput screens. We propose (1) development of the first on-chip ultra high-throughput whole- animal manipulation technologies for in vivo drug screens and genome-wide discovery of gene functions and interactions by complex on-chip behavioral and biochemical assay strategies; and (2) the first large-scale in vivo study of mechanisms of neural degeneration and regeneration following reproducible injuries using the proposed techniques. The proposed technologies and high-throughput assay strategies can significantly impact discovery of many molecular mechanisms using disease models of small animals.