Quantitative insights into age-associated DNA-repair inefficiency in single cells

ABSTRACT The double strand break (DSB) is a highly toxic form of DNA damage that is thought to be both a driver and consequence of age-related dysfunction. Although DSB repair is essential for a cell’s survival, little is known about how DSB repair mechanisms are affected by cellular age. Here we characterize the impact of cellular aging on the efficiency of single-strand annealing (SSA), a repair mechanism for DSBs occurring between direct repeats. Using a single-cell reporter of SSA repair, we measure SSA repair efficiency in young and old cells, and report a 23.4% decline in repair efficiency. This decline is not due to increased usage of non-homologous end joining (NHEJ). Instead, we identify increased G1-phase duration in old cells as a factor responsible for the decreased SSA repair efficiency. We further explore how SSA repair efficiency is affected by sequence heterology and find that heteroduplex rejection remains high in old cells. Our work provides novel quantitative insights into the links between cellular aging and DSB repair efficiency at single-cell resolution in replicatively aging cells.