title: Quantitating plant microRNA-mediated target repression using a dual-luciferase transient expression system.
abstract: microRNA (miRNA) mediated repression of target genes plays essential roles in a variety of functions in plants. An easy-to-use method that can effectively validate functional miRNA-target interactions in plants thus is of particular interest. Here, we describe an Agrobacterium tumefaciens-mediated in vivo assay utilizing a dual-luciferase reporter system. With this method, the strength of miRNA-mediated target repression can be quantified at both the mRNA (via qRT-PCR) and protein (via dual-luciferase assay) levels quickly and accurately.
authors.affiliation: Department of Biology, Pennsylvania State University, University Park, PA, 16802, USA.
authors.forename: Qikun
authors.initials: Q
authors.lastname: Liu
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Axtell
mesh_terms: D016960:Agrobacterium tumefaciens / Q000235:genetics; D015870:Gene Expression; D018506:Gene Expression Regulation, Plant; D017930:Genes, Reporter; D008156:Luciferases / Q000235:genetics*; D035683:MicroRNAs / Q000235:genetics*; D010944:Plants / Q000235:genetics*; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods
chemical_list: D035683:MicroRNAs; D012333:RNA, Messenger; D008156:Luciferases