title: Genome-Wide Study of mRNA Isoform Half-Lives.
abstract: In eukaryotes, RNA polymerase II-driven transcription and processing results in the formation of numerous mRNA 3' isoforms that for any given gene may differ from one another by as little as a single nucleotide. These 3' isoforms can vary in physical properties that may affect their function and stability. Here, we outline a systematic framework to measure individual mRNA 3' isoform half-lives on a genome-wide level in S. cerevisiae. Our approach utilizes the Anchor-Away system to sequester RNA polymerase II (Pol II) in the cytoplasm followed by direct single-molecule RNA sequencing to generate a highly detailed view of 3' isoform stability under most physiological conditions without many of the adverse effects associated with commonly used alternative approaches.
authors.affiliation: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, 240 Longwood Avenue, Boston, MA, 02115, USA. geisberg@hms.harvard.edu.
authors.forename: Joseph V
authors.initials: JV
authors.lastname: Geisberg
authors.affiliation: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, 240 Longwood Avenue, Boston, MA, 02115, USA.
authors.forename: Zarmik
authors.initials: Z
authors.lastname: Moqtaderi
mesh_terms: D016681:Genome, Fungal; D008967:Molecular Biology / Q000379:methods*; D059368:RNA Isoforms / Q000235:genetics*; D020871:RNA Stability / Q000235:genetics; D012331:RNA, Fungal / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics*; D012441:Saccharomyces cerevisiae; D017423:Sequence Analysis, RNA / Q000379:methods*; D014158:Transcription, Genetic
chemical_list: D059368:RNA Isoforms; D012331:RNA, Fungal; D012333:RNA, Messenger
keywords: Anchor-Away; RNA polymerase II; Saccharomyces cerevisiae transcription; mRNA half-life; mRNA isoform; mRNA stability