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.
In this proposal, we seek to develop a multiplexing microfluidics approach to study yeast budding and lifespan in a revolutionarily high throughput manner, to provide new understanding on aging. We propose to screen current and newly developed libraries of mutants to discover new genes and pathways that contribute to aging. We wish to discover key genes and pathways related to aging that may eventually contribute to increase the human lifespan/healthspan and improve the quality of life for the general public.
