Revolutionize Budding-Yeast-Based Aging Study by High-Throughput Lab-on-a-Chip Devices

 描述：衰老是许多导致死亡的疾病的主要危险因素。自从卡路里限制作为延长寿命的一种手段被发现后，许多研究工作都集中在揭示导致这种现象的分子机制上。遗憾的是，这些研究大多依赖于低吞吐量的方法，从而限制了进展。遗传上易驯化且生长相对较快的酿酒酵母模型为衰老研究提供了巨大的希望，因为许多衰老途径在所有真核生物中都是保守的。然而，它受到低通量方法、糟糕的显微技术以及缺乏基于单细胞的分析的太多限制。我们已经开发出一种创新和高效的微流控设备，它可以有效地捕获单亲母细胞并冲洗掉女儿，而不会在母细胞发芽时干扰它们。该设备与高通量显微镜方法相结合，提供了一种通过跟踪单个细胞来获取衰老过程信息的无与伦比的方法。通过这种创新的方法获得的数据的数量和类型是传统技术所不能实现的。我们已经使用这些设备收集了关于细胞形态和行为的新的和无与伦比的信息，因此，我们独一无二地准备通过在衰老基因的遗传筛选中使用这项技术来显著推进衰老研究。在这项提议中，我们寻求开发一种使用这种微流控设备的多路复用方法。我们建议筛选当前和新开发的突变体文库，包括ORF过表达文库和GFP融合文库，以发现与衰老有关的新基因和新途径。在完成这项拟议的工作后，我们将发现并报告与衰老相关的途径和蛋白质的新信息。这里提出的工作所确定的蛋白质将代表抗衰老策略开发的目标，这些策略可能会延长人类寿命并改变生活质量。

 DESCRIPTION: Aging is a primary risk factor for many diseases that lead to mortality. Upon the discovery of calorie restriction as a means to extend lifespan, much research effort has focused on uncovering the molecular mechanisms responsible for this phenomenon. Unfortunately, most of this research has depended upon low-throughput methods, thus limiting the progress. The genetically tractable and relatively fast-growing model, Saccharomyces cerevisiae, offers tremendous promise to contribute to aging research, as many of the aging pathways are conserved among all eukaryotes. However, it is too limited by low-throughput methods, poor microscopy techniques, and a lack of single cell-based assays. We have developed an innovative and highly effective microfluidic device that efficiently traps single mother cells and washes away daughters without disturbing the mother ones as they bud. This device, in combination with high-throughput microscopy methods, offers an unparalleled method to acquire information on the aging process by tracking single cells. The quantity and types of data acquired by this innovative method are impossible by the conventional techniques. We have already collected novel and unmatched information on cell morphology and behavior using these devices, and thus we are uniquely poised to markedly advance aging research by using this technique in genetic screens for aging genes. In this proposal, we seek to develop a multiplexing method, using this microfluidic device. We propose to screen current and newly developed libraries of mutants, including an ORF overexpression library and a GFP fusion library, to discover new genes and pathways that contribute to aging. Upon completion of this proposed work, we will have discovered and reported novel information on pathways and proteins associated with aging. The proteins identified by the work proposed here will represent targets for the development of anti-aging strategies that may increase the human lifespan and transform the quality of life.