title: 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,GFRA3is crucial for MDA-MB-231 cell growth, and its expression correlated with the overall survival of patients with breast cancer. Furthermore,GFRA3was 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.
authors.affiliation: Department of Systems BioMedicine, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University (TMDU), Bunkyo, Tokyo 113-8519, Japan.
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Ito
authors.affiliation: Department of Systems BioMedicine, National Research Institute for Child Health and Development, Setagaya, Tokyo 157-8535, Japan.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Inoue
authors.affiliation: Department of Systems BioMedicine, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University (TMDU), Bunkyo, Tokyo 113-8519, Japan.|Imperial College London, South Kensington, London SW7 2AZ, United Kingdom.
authors.forename: Timothy
authors.initials: T
authors.lastname: Seers
authors.affiliation: Department of Breast Surgery, Nagoya City University Graduate School of Medical Sciences, Nagoya, Aichi 467-8601, Japan.
authors.forename: Yukari
authors.initials: Y
authors.lastname: Hato
authors.affiliation: Department of Systems BioMedicine, National Research Institute for Child Health and Development, Setagaya, Tokyo 157-8535, Japan.
authors.forename: Arisa
authors.initials: A
authors.lastname: Igarashi
authors.affiliation: Department of Breast Surgery, Nagoya City University Graduate School of Medical Sciences, Nagoya, Aichi 467-8601, Japan.
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Toyama
authors.affiliation: Division of Biology, California Institute of Technology, Pasadena, CA 91125.
authors.forename: Konstantin D
authors.initials: KD
authors.lastname: Taganov
authors.affiliation: Department of Molecular and Cellular Biology, Beckman Research Institute, City of Hope, Duarte, CA 91010.
authors.forename: Mark P
authors.initials: MP
authors.lastname: Boldin
authors.affiliation: Department of Systems BioMedicine, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University (TMDU), Bunkyo, Tokyo 113-8519, Japan; asahara.syst@tmd.ac.jp.|Department of Systems BioMedicine, National Research Institute for Child Health and Development, Setagaya, Tokyo 157-8535, Japan.|Core Research for Evolutional Science and Technology (CREST), Japan Agency for Medical Research and Development (AMED), Tokyo 100-0004, Japan.|Department of Experimental Medicine, The Scripps Research Institute, San Diego, CA 92131.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Asahara
mesh_terms: D001943:Breast Neoplasms / Q000235:genetics* / Q000473:pathology; D045744:Cell Line, Tumor; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D015723:Gene Library; D017930:Genes, Reporter; D051097:Glial Cell Line-Derived Neurotrophic Factor Receptors / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D020365:RNA Helicases / Q000235:genetics; D015534:Trans-Activators / Q000235:genetics
chemical_list: C494993:GFRA3 protein, human; D051097:Glial Cell Line-Derived Neurotrophic Factor Receptors; C522425:MIRN34 microRNA, human; D035683:MicroRNAs; D015534:Trans-Activators; D020365:RNA Helicases; C101828:UPF1 protein, human
keywords: breast cancer; miR-34a; microRNA target screening; reporter library system