Ageing-related processes are largely conserved, with simple organisms remaining the main platform to discover and dissect new ageing-associated genes. Yeasts provide potent model systems to study cellular ageing owing their amenability to systematic functional assays under controlled conditions. Even with yeast cells, however, ageing assays can be laborious and resource-intensive. Here we present improved experimental and computational methods to study chronological lifespan in Schizosaccharomyces pombe. We decoded the barcodes for 3206 mutants of the latest gene-deletion library, enabling the parallel profiling of ~700 additional mutants compared to previous screens. We then applied a refined method of barcode sequencing (Bar-seq), addressing technical and statistical issues raised by persisting DNA in dead cells and sampling bottlenecks in aged cultures, to screen for mutants showing altered lifespan during stationary phase. This screen identified 341 long-lived mutants and 1246 short-lived mutants which point to many previously unknown ageing-associated genes, including 51 conserved but entirely uncharacterized genes. The ageing-associated genes showed coherent enrichments in processes also associated with human ageing, particularly with respect to ageing in non-proliferative brain cells. We also developed an automated colony-forming unit assay for chronological lifespan to facilitate medium- to high-throughput ageing studies by saving time and resources compared to the traditional assay. Results from the Bar-seq screen showed good agreement with this new assay, validating 33 genes not previously associated with cellular ageing. This study provides an effective methodological platform and identifies many new ageing-associated genes as a framework for analysing cellular ageing in yeast and beyond.

中文翻译：

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条形码测序和高通量分析的时间寿命发现裂变酵母中与衰老相关的基因

与衰老相关的过程在很大程度上得以保留，简单的生物仍然是发现和解剖新的与衰老相关的基因的主要平台。酵母由于能够在受控条件下进行系统功能分析，因此提供了用于研究细胞衰老的有效模型系统。然而，即使使用酵母细胞，老化测定也可能是费力且耗费资源的。在这里，我们提出了改进的实验和计算方法来研究粟酒裂殖酵母的时间顺序寿命。我们对最新基因缺失文库的3206个突变体的条形码进行了解码，与以前的筛选相比，能够并行分析约700个其他突变体。然后，我们应用了一种改进的条形码测序方法（Bar-seq），解决了将DNA保留在死细胞中以及老化培养物中的采样瓶颈所引起的技术和统计问题，以筛选在固定阶段显示寿命改变的突变体。此筛选确定了341个长寿突变体和1246个短寿突变体，这些突变体指向许多以前未知的衰老相关基因，包括51个保守但完全未表征的基因。与衰老相关的基因在与人类衰老相关的过程中表现出一致的富集，特别是在非增生性脑细胞中的衰老方面。我们还针对时间寿命开发了一种自动菌落形成单位测定法，与传统测定法相比，它通过节省时间和资源来促进中至高通量的衰老研究。Bar-seq筛查结果与这项新检测方法显示出良好的一致性，验证了以前与细胞衰老无关的33个基因。这项研究提供了一个有效的方法学平台，并确定了许多新的与衰老相关的基因，作为分析酵母及其他细胞衰老的框架。