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Identification of targets of tumor suppressor microRNA-34a using a reporter library system.


ABSTRACT: miRNAs play critical roles in various biological processes by targeting specific mRNAs. Current approaches to identifying miRNA targets are insufficient for elucidation of a miRNA regulatory network. Here, we created a cell-based screening system using a luciferase reporter library composed of 4,891 full-length cDNAs, each of which was integrated into the 3' UTR of a luciferase gene. Using this reporter library system, we conducted a screening for targets of miR-34a, a tumor-suppressor miRNA. We identified both previously characterized and previously uncharacterized targets. miR-34a overexpression in MDA-MB-231 breast cancer cells repressed the expression of these previously unrecognized targets. Among these targets, GFRA3 is crucial for MDA-MB-231 cell growth, and its expression correlated with the overall survival of patients with breast cancer. Furthermore, GFRA3 was found to be directly regulated by miR-34a via its coding region. These data show that this system is useful for elucidating miRNA functions and networks.

SUBMITTER: Ito Y 

PROVIDER: S-EPMC5393199 | biostudies-literature | 2017 Apr

REPOSITORIES: biostudies-literature

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Identification of targets of tumor suppressor microRNA-34a using a reporter library system.

Ito Yoshiaki Y   Inoue Atsushi A   Seers Timothy T   Hato Yukari Y   Igarashi Arisa A   Toyama Tatsuya T   Taganov Konstantin D KD   Boldin Mark P MP   Asahara Hiroshi H  

Proceedings of the National Academy of Sciences of the United States of America 20170329 15


miRNAs play critical roles in various biological processes by targeting specific mRNAs. Current approaches to identifying miRNA targets are insufficient for elucidation of a miRNA regulatory network. Here, we created a cell-based screening system using a luciferase reporter library composed of 4,891 full-length cDNAs, each of which was integrated into the 3' UTR of a luciferase gene. Using this reporter library system, we conducted a screening for targets of miR-34a, a tumor-suppressor miRNA. We  ...[more]

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