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

被引:32
作者
Lataniotis, Lazaros [1 ]
Albrecht, Andreas [2 ]
Kok, Fatma O. [3 ]
Monfries, Clinton A. L. [4 ]
Benedetti, Lorena [5 ]
Lawson, Nathan D. [3 ]
Hughes, Simon M. [4 ]
Steinhofel, Kathleen [6 ]
Mayr, Manuel [1 ]
Zampetaki, Anna [1 ]
机构
[1] Kings Coll London, Kings British Heart Fdn Ctr, London, England
[2] Middlesex Univ, Sch Sci & Technol, London, England
[3] Univ Massachusetts, Sch Med, Dept Mol Cell & Canc Biol, Worcester, MA USA
[4] Kings Coll London, Randall Div, London, England
[5] Kings Coll London, Div Canc Studies, London, England
[6] Kings Coll London, Informat Dept, London, England
来源
SCIENTIFIC REPORTS | 2017年 / 7卷
基金
英国惠康基金; 英国医学研究理事会;
关键词
FAMILY; GENE; CRISPR-CAS9; KNOCKOUT; CELLS; DETERMINANTS; TRANSCRIPTS; GENOMICS; ROLES;
D O I
10.1038/s41598-017-09268-0
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
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 similar to 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 similar to 195 that may affect its processing. In cluster miR-143 similar to 145 our results imply a feed-forward regulation, although structural changes cannot be ruled out. Furthermore, in the miR-179 similar to and miR-106 similar to 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.
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页数:14
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