Scientific Reports (Aug 2017)

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

  • Lazaros Lataniotis,
  • Andreas Albrecht,
  • Fatma O. Kok,
  • Clinton A. L. Monfries,
  • Lorena Benedetti,
  • Nathan D. Lawson,
  • Simon M. Hughes,
  • Kathleen Steinhofel,
  • Manuel Mayr,
  • Anna Zampetaki

DOI
https://doi.org/10.1038/s41598-017-09268-0
Journal volume & issue
Vol. 7, no. 1
pp. 1 – 14

Abstract

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Abstract 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.