CRISPR/Cas9 editing reveals novel mechanisms of clustered microRNA regulation and function

• Published in: Scientific reports Vol. 7; no. 1; pp. 8585 - 14
• Main Authors: Lataniotis, Lazaros, Albrecht, Andreas, Kok, Fatma O., Monfries, Clinton A. L., Benedetti, Lorena, Lawson, Nathan D., Hughes, Simon M., Steinhofel, Kathleen, Mayr, Manuel, Zampetaki, Anna
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 17.08.2017; Nature Publishing Group; Nature Portfolio
• Subjects: 38; 38/109; 38/22; 38/77; 38/90; 45; 45/41; 631/1647/1511; 631/337/384/331; Animals; Antibiotics; Cancer; Cells, Cultured; Computer applications; Computer Simulation; CRISPR; CRISPR-Cas Systems; Gene Editing; Gene expression; Gene Expression Profiling; Gene loci; Genome editing; HEK293 Cells; Humanities and Social Sciences; Humans; Mice; MicroRNAs; MicroRNAs - genetics; MicroRNAs - metabolism; miRNA; multidisciplinary; Multigene Family; Muscle, Smooth, Vascular - cytology; Science; Science (multidisciplinary)
• ISSN: 2045-2322; 2045-2322
• 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-497~195 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-497~195 that may affect its processing. In cluster miR-143~145 our results imply a feed-forward regulation, although structural changes cannot be ruled out. Furthermore, in the miR-17~92 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.