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

Ito, Yoshiaki and Inoue, Atsushi and Seers, Timothy and Hato, Yukari and Igarashi, Arisa and Toyama, Tatsuya and Taganov, Konstantin D. and Boldin, Mark P. and Asahara, Hiroshi (2017) Identification of targets of tumor suppressor microRNA-34a using a reporter library system. Proceedings of the National Academy of Sciences of the United States of America, 114 (15). pp. 3927-3932. ISSN 0027-8424. https://resolver.caltech.edu/CaltechAUTHORS:20170329-152704022

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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.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1073/pnas.1620019114DOIArticle
http://www.pnas.org/content/114/15/3927PublisherArticle
http://www.pnas.org/content/114/15/3927/suppl/DCSupplementalPublisherSupporting Information
ORCID:
AuthorORCID
Ito, Yoshiaki0000-0002-9037-1184
Igarashi, Arisa0000-0001-7206-3105
Boldin, Mark P.0000-0003-4593-0669
Additional Information:© 2017 National Academy of Sciences. Edited by Tak W. Mak, The Campbell Family Institute for Breast Cancer Research at Princess Margaret Cancer Centre, University Health Network, Toronto, Canada, and approved March 9, 2017 (received for review December 8, 2016). Published online before print March 29, 2017. We thank Drs. Akio Yamashita, Masato Yano, Tomoaki Tanaka, and Kazuhiko Kuwahara for helpful discussions and Mayuko Yoda, Yukinari Osaka, Nobuhiro Yamamoto, and Ayaka Ichise for technical assistance. This work was supported by Core Research for the Evolutionary Science and Technology (CREST) funding from the Japan Science and Technology Agency; CREST funding from the Japan Agency for Medical Research and Development; Grants-in-Aid for Scientific Research (KAKENHI) Grants 26113008, 15H02560, 15K15544, 15K15026, and 25860139 from the Japan Society for the Promotion of Science; NIH Grants AR050631 and AR065379; the Naito Foundation; and a Bristol-Myers K.K. RA Clinical Investigation Grant. Author contributions: Y.I., A. Inoue, and H.A. designed research; Y.I., A. Inoue, T.S., Y.H., A. Igarashi, and T.T. performed research; Y.I., A. Inoue, and H.A. contributed new reagents/analytic tools; Y.I., A. Inoue, T.S., Y.H., T.T., K.D.T., M.P.B., and H.A. analyzed data; and Y.I. and H.A. wrote the paper. The authors declare no conflict of interest. This article is a PNAS Direct Submission. This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10.1073/pnas.1620019114/-/DCSupplemental.
Funders:
Funding AgencyGrant Number
Japan Science and Technology Agency (JST)UNSPECIFIED
Japan Agency for Medical Research and DevelopmentUNSPECIFIED
Japan Society for the Promotion of Science (JSPS)26113008
Japan Society for the Promotion of Science (JSPS)15H02560
Japan Society for the Promotion of Science (JSPS)15K15544
Japan Society for the Promotion of Science (JSPS)15K15026
Japan Society for the Promotion of Science (JSPS)25860139
NIHAR050631
NIHAR065379
Naito FoundationUNSPECIFIED
Bristol-MyersUNSPECIFIED
Subject Keywords:microRNA target screening; miR-34a; reporter library system; breast cancer
Issue or Number:15
Record Number:CaltechAUTHORS:20170329-152704022
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20170329-152704022
Official Citation:Yoshiaki Ito, Atsushi Inoue, Timothy Seers, Yukari Hato, Arisa Igarashi, Tatsuya Toyama, Konstantin D. Taganov, Mark P. Boldin, and Hiroshi Asahara Identification of targets of tumor suppressor microRNA-34a using a reporter library system PNAS 2017 114 (15) 3927-3932; published ahead of print March 29, 2017, doi:10.1073/pnas.1620019114
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:75522
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:29 Mar 2017 22:54
Last Modified:03 Oct 2019 16:51

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