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


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.

  • 著者: Yoshiaki Ito, Atsushi Inoue, Timothy Seers, Yukari Hato, Arisa Igarashi, Tatsuya Toyama, Konstantin D. Taganov, Mark P. Boldin, and Hiroshi Asahara
  • 掲載誌: Proc Natl Acad Sci U S A



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