Microfluids-enabled Quantitative Systems Biology Approaches to Construct Gene Networks Regulating Yeast Lifespan and Genetic Noise Dynamics During Aging

摘要 老龄化是人类慢性发病率和死亡率的主要原因。针对关键蛋白质的干预措施 因此，在细胞和生物层面延长健康寿命是可取的。延伸的基因变化 模式生物的寿命通常会推迟与年龄相关的疾病的发生，并显示出进化上的保守性 在高等生物体中。细胞通常使用基因网络来协调它们对刺激的反应，因此 细胞老化的动力学也可能在基因网络水平上受到控制。然而，人们对此知之甚少 这种衰老网络的组成既涉及到基因，也涉及到它们之间的关系 网络。通过在微流控设备中对酵母复制寿命进行高通量测量，这 该提案旨在确定并在功能上划分新型衰老模块的遗传成分 基于上位性相互作用的强度。此外，它还旨在阐明这些网络的影响 模块在老化过程中降低噪音，这是单电池老化的一种紧急特性。的关键组件 老化模块以及来自染色质重塑网络的基因将从酵母基因组中删除 由此产生的遗传噪声动力学的变化将使用典型的GAL1的活动来测量 启动子在单个酵母细胞中作为模型。最后，组成基因的关键基因的表达强度变化 老化模块将在细胞老化过程中进行实验跟踪，目的是探索衰老是如何改变的 网络节点的可塑性。该项目的成功完成将在复制之间提供新的联系 新发现的网络模块背景下的寿命和基因表达噪声的集体表型 实时控制单个酵母细胞的老化情况。

ABSTRACT Aging is the primary cause of chronic human morbidity and mortality. Interventions targeting key proteins that extend healthy lifespan at the cellular and organismal level are therefore desirable. Genetic changes that extend lifespan in model organisms often delay the onset of age-related morbidity and exhibit evolutionary conservation in higher organisms. Cells typically use gene networks to coordinate their response to stimuli, and thus the dynamics of cellular aging is also likely governed at the gene network level. However, little is known about the composition of such an aging network both in terms of the genes involved and their relationship within the network. By performing high-throughput measurements of yeast replicative lifespan in a microfluidic device, this proposal aims to identify and functionally compartmentalize the genetic components of novel aging modules based on the strength of epistatic interactions. Furthermore, it aims to elucidate the impact of these network modules on noise reduction during aging, an emergent property of single-cell aging. Key components of the aging modules as well as genes from a chromatin-remodeling network will be deleted from the yeast genome and the resulting changes in genetic noise dynamics will be measured using the activity of the canonical GAL1 promoter as a model in single yeast cells. Finally, expression strength changes of the key genes composing the aging modules will be followed experimentally during cellular aging with the goal of exploring how aging alters the plasticity of network nodes. Successful completion of this project will provide novel links between replicative lifespan and the collective phenotype of gene expression noise in the context of newly identified network modules governing how single yeast cells age in real time.