title: Methods for studying IRES-mediated translation of positive-strand RNA viruses.
abstract: Internal ribosome entry sites are RNA elements that mediate translation in a cap-independent manner. A subset of positive strand RNA viruses utilize an IRES mechanism as a viral strategy to ensure efficient viral protein synthesis. IRES elements vary in sequence, structure, and factor requirements between virus families. Here, we describe methods to determine IRES activity and approaches to study the regulation and function of IRES-mediated translation both in vitro and in vivo. Finally, we describe a new IRES-directed reporter system which exploits the 2A 'self-cleavage' or 'stop-go' peptide for optimal detection of IRES activity.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, BC, Canada V6T 1Z3.
authors.forename: Qing S
authors.initials: QS
authors.lastname: Wang
authors.forename: Hilda H T
authors.initials: HH
authors.lastname: Au
authors.forename: Eric
authors.initials: E
authors.lastname: Jan
mesh_terms: D000818:Animals; D001483:Base Sequence; D005821:Genetic Techniques; D006801:Humans; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis / Q000235:genetics*; D012328:RNA Viruses / Q000235:genetics*; D012270:Ribosomes / Q000235:genetics* / Q000821:virology*