title: Effective experimental validation of miRNA targets using an improved linker reporter assay.
abstract: miRNAs are small, non-coding RNAs that play critical roles in various cellular processes. Although there are several algorithms that can predict the potential candidate genes that are regulated by a miRNA, these algorithms require further experimental validation in order to demonstrate genuine targets of miRNAs. Moreover, most algorithms predict hundreds to thousands of putative target genes for each miRNA, and it is difficult to validate all candidates using the whole 3'-untranslated region (UTR) reporter assay. We report a fast, simple and efficient experimental approach to screening miRNA candidate targets using a 3'-UTR linker assay. Critically, the linker has only a short miRNA regulatory element sequence of approximately 22 base pairs in length and can provide a benefit for screening strong miRNA candidates for further validation using the whole 3'-UTR sequence. Our technique will provide a simplified platform for the high-throughput screening of miRNA target gene validation.
authors.affiliation: Soonchunhyang Institute of Med-bioscience (SIMS),Sunchunhyang University,Chonan-Si,336-745,Republic of Korea.
authors.forename: Cheolwon
authors.initials: C
authors.lastname: Choi
authors.affiliation: Great Oak High School,Temecula,CA,USA.
authors.forename: James
authors.initials: J
authors.lastname: Han
authors.affiliation: Soonchunhyang Institute of Med-bioscience (SIMS),Sunchunhyang University,Chonan-Si,336-745,Republic of Korea.
authors.forename: Nguyen Thi
authors.initials: NT
authors.lastname: Thao Tran
authors.affiliation: Soonchunhyang Institute of Med-bioscience (SIMS),Sunchunhyang University,Chonan-Si,336-745,Republic of Korea.
authors.forename: Seulgi
authors.initials: S
authors.lastname: Yoon
authors.affiliation: Soonchunhyang Institute of Med-bioscience (SIMS),Sunchunhyang University,Chonan-Si,336-745,Republic of Korea.
authors.forename: Goeun
authors.initials: G
authors.lastname: Kim
authors.affiliation: Soonchunhyang Institute of Med-bioscience (SIMS),Sunchunhyang University,Chonan-Si,336-745,Republic of Korea.
authors.forename: Sujung
authors.initials: S
authors.lastname: Song
authors.affiliation: Soonchunhyang Institute of Med-bioscience (SIMS),Sunchunhyang University,Chonan-Si,336-745,Republic of Korea.
authors.forename: Youngjo
authors.initials: Y
authors.lastname: Kim
authors.affiliation: Soonchunhyang Institute of Med-bioscience (SIMS),Sunchunhyang University,Chonan-Si,336-745,Republic of Korea.
authors.forename: Seongho
authors.initials: S
authors.lastname: Ryu
mesh_terms: D020413:3' Untranslated Regions / Q000235:genetics*; D001483:Base Sequence; D014408:Biomarkers, Tumor / Q000235:genetics / Q000378:metabolism*; D015153:Blotting, Western; D019295:Computational Biology; D015972:Gene Expression Regulation, Neoplastic; D057809:HEK293 Cells; D006801:Humans; D008156:Luciferases / Q000378:metabolism*; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D009369:Neoplasms / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*
chemical_list: D020413:3' Untranslated Regions; D014408:Biomarkers, Tumor; D035683:MicroRNAs; D012333:RNA, Messenger; D008156:Luciferases