title: In Vitro Reversible Translation Control Using γPNA Probes.
abstract: On-demand regulation of gene expression in living cells is a central goal of chemical biology and antisense therapeutic development. While significant advances have allowed regulatory modulation through inserted genetic elements, on-demand control of the expression/translation state of a given native gene by complementary sequence interactions remains a technical challenge. Toward this objective, we demonstrate the reversible suppression of a luciferase gene in cell-free translation using Watson-Crick base pairing between the mRNA and a complementary γ-modified peptide nucleic acid (γPNA) sequence with a noncomplementary toehold. Exploiting the favorable thermodynamics of γPNA-γPNA interactions, the antisense sequence can be removed by hybridization of a second, fully complementary γPNA, through a strand displacement reaction, allowing translation to proceed. Complementary RNA is also shown to displace the bound antisense γPNA, opening up possibilities of in vivo regulation by native gene expression.
authors.affiliation: †Department of Chemistry, ‡Department of Biological Sciences, and §Center for Nucleic Acids Science and Technology, Carnegie Mellon University, 4400 Fifth Avenue, Pittsburgh, Pennsylvania 15213, United States.
authors.forename: Taylor D
authors.initials: TD
authors.lastname: Canady
authors.affiliation: †Department of Chemistry, ‡Department of Biological Sciences, and §Center for Nucleic Acids Science and Technology, Carnegie Mellon University, 4400 Fifth Avenue, Pittsburgh, Pennsylvania 15213, United States.
authors.forename: Cheryl A
authors.initials: CA
authors.lastname: Telmer
authors.affiliation: †Department of Chemistry, ‡Department of Biological Sciences, and §Center for Nucleic Acids Science and Technology, Carnegie Mellon University, 4400 Fifth Avenue, Pittsburgh, Pennsylvania 15213, United States.
authors.forename: Stanley N
authors.initials: SN
authors.lastname: Oyaghire
authors.affiliation: †Department of Chemistry, ‡Department of Biological Sciences, and §Center for Nucleic Acids Science and Technology, Carnegie Mellon University, 4400 Fifth Avenue, Pittsburgh, Pennsylvania 15213, United States.
authors.forename: Bruce A
authors.initials: BA
authors.lastname: Armitage
authors.affiliation: †Department of Chemistry, ‡Department of Biological Sciences, and §Center for Nucleic Acids Science and Technology, Carnegie Mellon University, 4400 Fifth Avenue, Pittsburgh, Pennsylvania 15213, United States.
authors.forename: Marcel P
authors.initials: MP
authors.lastname: Bruchez
mesh_terms: D000818:Animals; D020029:Base Pairing; D002474:Cell-Free System; D008156:Luciferases / Q000235:genetics; D009693:Nucleic Acid Hybridization; D015341:Nucleic Acid Probes / Q000737:chemistry*; D020135:Peptide Nucleic Acids / Q000737:chemistry* / Q000235:genetics; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000737:chemistry / Q000235:genetics; D011817:Rabbits; D012156:Reticulocytes / Q000737:chemistry; D013816:Thermodynamics
chemical_list: D015341:Nucleic Acid Probes; D020135:Peptide Nucleic Acids; D012333:RNA, Messenger; D008156:Luciferases