Institute of Functional Epigenetics, Helmholtz Zentrum München, 85764 Neuherberg, Germany.
Institute of Functional Epigenetics, Helmholtz Zentrum München, 85764 Neuherberg, Germany; Instituto de Biotecnologia, National Autonomous University of Mexico, Cuernavaca 62210, Mexico.
Cell Rep. 2022 Nov 15;41(7):111656. doi: 10.1016/j.celrep.2022.111656.
Asymmetric inheritance of cellular content through cell division plays an important role in cell viability and fitness. The dynamics of RNA segregation are so far largely unaddressed. This is partly due to a lack of approaches to follow RNAs over multiple cellular divisions. Here, we establish an approach to quantify RNA dynamics in single cells across several generations in a microfluidics device by tagging RNAs with the diSpinach aptamer. Using S. cerevisiae as a model, we quantitatively characterize intracellular RNA transport from mothers into their buds. Our results suggest that, at cytokinesis, ENO2 diSpinach RNA is preferentially distributed to daughters. This asymmetric RNA segregation depends on the lifespan regulator Sir2 and decreases with increasing replicative age of mothers but does not result from increasing cell size during aging. Overall, our approach opens more opportunities to study RNA dynamics and inheritance in live budding yeast at the single-cell level.
通过细胞分裂实现的细胞内容物的非对称遗传对细胞活力和适应性起着重要作用。RNA 分离的动力学至今在很大程度上尚未得到解决。这部分是由于缺乏在多个细胞分裂过程中跟踪 RNA 的方法。在这里，我们通过将 diSpinach 适体标记 RNA，在微流控设备中建立了一种在多个世代中定量研究单个细胞中 RNA 动力学的方法。我们使用 S. cerevisiae 作为模型，定量描述了从母细胞到其芽细胞的细胞内 RNA 转运。我们的结果表明，在胞质分裂时，ENO2 diSpinach RNA 优先分配给子细胞。这种不对称的 RNA 分离依赖于寿命调节剂 Sir2，并且随着母细胞复制年龄的增加而减少，但不是由于衰老过程中细胞尺寸的增加所致。总的来说，我们的方法为在单细胞水平上研究活 budding yeast 中的 RNA 动力学和遗传提供了更多机会。
