title: Gain-of-function reporters for analysis of mRNA 3'-end formation: Design and optimization.
abstract: The concept of mRNA 3'-end formation as a static, minimally regulated housekeeping process has undergone a paradigm shift. Many recent studies have shown that accurate and efficient 3'-end formation of mRNA is highly regulated and that dysregulation of this process is a hallmark of several diseases. While there are many global analysis methods for monitoring altered mRNA processing, methods for investigating specific RNA 3'-end processing events in cells have not significantly changed. Here we describe a facile gain-of-function cellular reporter for the analysis of mRNA 3'-end formation as an alternative to approaches that are technically challenging or use radioactivity. We also offer suggestions for optimization of our approach and enhancement of its reproducibility.
authors.affiliation: Department of Biochemistry and Molecular Biology, The University of Texas Medical School at Houston, Houston, TX.|Graduate Program in Biochemistry and Molecular Biology, The University of Texas Graduate School of Biomedical Sciences, Houston, TX.|Department of Biochemistry and Molecular Biology, The University of Texas Medical Branch, Galveston, TX.
authors.forename: Natoya
authors.initials: N
authors.lastname: Peart
authors.affiliation: Department of Biochemistry and Molecular Biology, The University of Texas Medical School at Houston, Houston, TX.|Graduate Program in Biochemistry and Molecular Biology, The University of Texas Graduate School of Biomedical Sciences, Houston, TX.|Department of Biochemistry and Molecular Biology, The University of Texas Medical Branch, Galveston, TX.
authors.forename: Eric J
authors.initials: EJ
authors.lastname: Wagner
mesh_terms: D017930:Genes, Reporter / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000235:genetics*; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D012313:RNA / Q000235:genetics*; D026721:RNA 3' End Processing / Q000235:genetics*; D017423:Sequence Analysis, RNA / Q000379:methods*; D059467:Transcriptome / Q000235:genetics*
chemical_list: D049452:Green Fluorescent Proteins; D012313:RNA
keywords: 3′-end processing; gain-of-function; reporter assay