King's British Heart Foundation Centre, King's College London, London, UK.
Middlesex University, School of Science and Technology, London, UK.
Sci Rep. 2017 Aug 17;7(1):8585. doi: 10.1038/s41598-017-09268-0.
MicroRNAs (miRNAs) are important regulators of diverse physiological and pathophysiological processes. MiRNA families and clusters are two key features in miRNA biology. Here we explore the use of CRISPR/Cas9 as a powerful tool to delineate the function and regulation of miRNA families and clusters. We focused on four miRNA clusters composed of miRNA members of the same family, homo-clusters or different families, hetero-clusters. Our results highlight different regulatory mechanisms in miRNA cluster expression. In the case of the miR-497195 cluster, editing of miR-195 led to a significant decrease in the expression of the other miRNA in the cluster, miR-497a. Although no gene editing was detected in the miR-497a genomic locus, computational simulation revealed alteration in the three dimensional structure of the pri-miR-497195 that may affect its processing. In cluster miR-143145 our results imply a feed-forward regulation, although structural changes cannot be ruled out. Furthermore, in the miR-1792 and miR-106~25 clusters no interdependency in miRNA expression was observed. Our findings suggest that CRISPR/Cas9 is a powerful gene editing tool that can uncover novel mechanisms of clustered miRNA regulation and function.
微小 RNA（miRNA）是调节多种生理和病理生理过程的重要调控因子。miRNA 家族和簇是 miRNA 生物学的两个关键特征。在这里，我们探讨了使用 CRISPR/Cas9 作为一种强大的工具来描绘 miRNA 家族和簇的功能和调节。我们专注于四个由同一家族成员 miRNA 组成的 miRNA 簇、同家族或不同家族的 miRNA 簇、异家族 miRNA 簇。我们的结果强调了 miRNA 簇表达中的不同调节机制。在 miR-497195 簇的情况下，miR-195 的编辑导致该簇中另一个 miRNA miR-497a 的表达显著下降。尽管在 miR-497a 基因组位置未检测到基因编辑，但计算模拟显示 pri-miR-497195 的三维结构发生改变，可能影响其加工。在 miR-143145 簇中，我们的结果表明存在正反馈调节，尽管不能排除结构变化。此外，在 miR-1792 和 miR-106~25 簇中，miRNA 表达没有相互依赖性。我们的发现表明，CRISPR/Cas9 是一种强大的基因编辑工具，可以揭示簇状 miRNA 调节和功能的新机制。
