Telethon Institute of Genetics and Medicine, Pozzuoli, 80078 Naples, Italy.
The Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, Center for Reproductive Sciences, University of California, San Francisco, San Francisco, California 94143, USA.
Nat Commun. 2017 Jan 19;8:14126. doi: 10.1038/ncomms14126.
MicroRNAs act posttranscriptionally to suppress multiple target genes within a cell population. To what extent this multi-target suppression occurs in individual cells and how it impacts transcriptional heterogeneity and gene co-expression remains unknown. Here we used single-cell sequencing combined with introduction of individual microRNAs. miR-294 and let-7c were introduced into otherwise microRNA-deficient Dgcr8 knockout mouse embryonic stem cells. Both microRNAs induce suppression and correlated expression of their respective gene targets. The two microRNAs had opposing effects on transcriptional heterogeneity within the cell population, with let-7c increasing and miR-294 decreasing the heterogeneity between cells. Furthermore, let-7c promotes, whereas miR-294 suppresses, the phasing of cell cycle genes. These results show at the individual cell level how a microRNA simultaneously has impacts on its many targets and how that in turn can influence a population of cells. The findings have important implications in the understanding of how microRNAs influence the co-expression of genes and pathways, and thus ultimately cell fate.
微小 RNA 通过转录后作用抑制细胞群体内的多个靶基因。在单个细胞中，这种多靶抑制在何种程度上发生，以及它如何影响转录异质性和基因共表达，目前尚不清楚。在这里，我们使用单细胞测序结合引入单个 microRNA 的方法。miR-294 和 let-7c 被引入到 otherwise microRNA 缺乏的 Dgcr8 敲除小鼠胚胎干细胞中。这两种 microRNA 都诱导它们各自的基因靶标的抑制和相关表达。这两种 microRNA 对细胞群体内的转录异质性有相反的影响，let-7c 增加而 miR-294 减少细胞之间的异质性。此外，let-7c 促进细胞周期基因的相位，而 miR-294 抑制。这些结果显示了 microRNA 如何在单个细胞水平上同时对其许多靶基因产生影响，以及这反过来又如何影响细胞群体。这些发现对于理解 microRNA 如何影响基因和途径的共表达，以及最终影响细胞命运具有重要意义。
