title: Use of the pBUTR Reporter System for Scalable Analysis of 3' UTR-Mediated Gene Regulation.
abstract: Posttranscriptional control of mRNA subcellular localization, stability, and translation is a central aspect of gene regulation and expression. Much of this control is mediated via recognition of a given mRNA transcript's 3' untranslated region (UTR) by microRNAs and RNA-binding proteins. Here we describe how a novel, scalable piggyBac-based vector, pBUTR, can be utilized for analysis of 3' UTR-mediated posttranscriptional gene regulation (PTGR) both in vitro and in vivo. This vector is specifically designed to express a selection marker, a control reporter, and an experimental reporter from three independent transcription units. Expression of spliced reporter transcripts from medium-copy non-viral promoter elements circumvents several potential confounding factors associated with saturation and stability, while stable integration of these reporter and selection elements in the context of a DNA transposon facilitates experimental reproducibility.
authors.affiliation: Department of Molecular Physiology and Biophysics and Dan L. Duncan Cancer Center, Baylor College of Medicine, Houston, TX, 77030, USA.
authors.forename: Arindam
authors.initials: A
authors.lastname: Chaudhury
authors.affiliation: Department of Molecular Physiology and Biophysics and Dan L. Duncan Cancer Center, Baylor College of Medicine, Houston, TX, 77030, USA. neilson@bcm.edu.
authors.forename: Joel R
authors.initials: JR
authors.lastname: Neilson
mesh_terms: D020413:3' Untranslated Regions / Q000235:genetics; D002460:Cell Line; D005434:Flow Cytometry / Q000379:methods*; D020869:Gene Expression Profiling / Q000379:methods*; D005786:Gene Expression Regulation / Q000235:genetics; D017930:Genes, Reporter / Q000235:genetics; D006801:Humans; D014176:Protein Biosynthesis / Q000235:genetics; D012323:RNA Processing, Post-Transcriptional / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics*; D016601:RNA-Binding Proteins / Q000096:biosynthesis / Q000235:genetics; D014162:Transfection
chemical_list: D020413:3' Untranslated Regions; D012333:RNA, Messenger; D016601:RNA-Binding Proteins
keywords: 3′-UTR; PTGR; Posttranscriptional gene regulation; RNA-binding proteins; Reporter; mRNA stability; miRNA sensors; pBUTR; piggyBac