title: Cellular microRNA sensors based on luciferase reporters.
abstract: Recently, microRNAs (miRNAs) have been linked to a variety of human diseases including cancer and viral infections. Small molecule modifiers of miRNAs could represent new therapeutic agents and be used as tools for elucidating the biological roles of miRNAs. In order to identify small molecule modifiers of miRNAs, functional assays for specific miRNAs must be developed and optimized. Here, we report the construction of a luciferase reporter assay for miRNA miR-122 function and the development of a stable Huh7 cell line that can be used for high-throughput screening of small molecule miR-122 inhibitors. The steps described here can be applied not only to Huh7 cells and miR-122 but also to virtually any cell line and miRNA combination.
authors.affiliation: Department of Chemistry, North Carolina State University, Raleigh, NC, USA.
authors.forename: Colleen M
authors.initials: CM
authors.lastname: Connelly
authors.forename: Alexander
authors.initials: A
authors.lastname: Deiters
mesh_terms: D015374:Biosensing Techniques / Q000379:methods*; D045744:Cell Line, Tumor; D017930:Genes, Reporter / Q000235:genetics*; D006801:Humans; D049410:Luciferases, Renilla / Q000235:genetics*; D035683:MicroRNAs / Q000032:analysis* / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D014162:Transfection
chemical_list: C531907:MIRN122 microRNA, human; D035683:MicroRNAs; D049410:Luciferases, Renilla