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

TY - JOUR

T1 - CRISPR/Cas9 editing reveals novel mechanisms of clustered microRNA regulation and function

AU - Lataniotis, Lazaros

AU - Albrecht, Andreas

AU - Kok, Fatma O

AU - Monfries, Clinton A L

AU - Benedetti, Lorena

AU - Lawson, Nathan D

AU - Hughes, Simon

AU - Steinhofel, Kathleen

AU - Mayr, Manuel

AU - Zampetaki, Anna

PY - 2017/8/17

Y1 - 2017/8/17

N2 - 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.

AB - 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.

KW - Journal Article

U2 - 10.1038/s41598-017-09268-0

DO - 10.1038/s41598-017-09268-0

M3 - Article

C2 - 28819307

SN - 2045-2322

VL - 7

SP - 8585

JO - Scientific Reports

JF - Scientific Reports

IS - 1

ER -