title: RNA G-Quadruplex Invasion and Translation Inhibition by Antisense γ-Peptide Nucleic Acid Oligomers.
abstract: We have examined the abilities of three complementary γ-peptide nucleic acid (γPNA) oligomers to invade an RNA G-quadruplex and potently inhibit translation of a luciferase reporter transcript containing the quadruplex-forming sequence (QFS) within its 5'-untranslated region. All three γPNA oligomers bind with low nanomolar affinities to an RNA oligonucleotide containing the QFS. However, while all probes inhibit translation with low to midnanomolar IC50 values, the γPNA designed to hybridize to the first two G-tracts of the QFS and adjacent 5'-overhanging nucleotides was 5-6 times more potent than probes directed to either the 3'-end or internal regions of the target at 37 °C. This position-dependent effect was eliminated after the probes and target were preincubated at an elevated temperature prior to translation, demonstrating that kinetic effects exert significant control over quadruplex invasion and translation inhibition. We also found that antisense γPNAs exhibited similarly potent effects against luciferase reporter transcripts bearing QFS motifs having G2, G3, or G4 tracts. Finally, our results indicate that γPNA oligomers exhibit selectivity and/or potency higher than those of other antisense molecules such as standard PNA and 2'-OMe RNA previously reported to target G-quadruplexes in RNA.
authors.affiliation: Department of Chemistry and Center for Nucleic Acids Science and Technology, Carnegie Mellon University , 4400 Fifth Avenue, Pittsburgh, Pennsylvania 15213-3890, United States.
authors.forename: Stanley N
authors.initials: SN
authors.lastname: Oyaghire
authors.affiliation: Department of Chemistry and Center for Nucleic Acids Science and Technology, Carnegie Mellon University , 4400 Fifth Avenue, Pittsburgh, Pennsylvania 15213-3890, United States.
authors.forename: Collin J
authors.initials: CJ
authors.lastname: Cherubim
authors.affiliation: Department of Chemistry and Center for Nucleic Acids Science and Technology, Carnegie Mellon University , 4400 Fifth Avenue, Pittsburgh, Pennsylvania 15213-3890, United States.
authors.forename: Cheryl A
authors.initials: CA
authors.lastname: Telmer
authors.affiliation: Department of Chemistry and Center for Nucleic Acids Science and Technology, Carnegie Mellon University , 4400 Fifth Avenue, Pittsburgh, Pennsylvania 15213-3890, United States.
authors.forename: Joe A
authors.initials: JA
authors.lastname: Martinez
authors.affiliation: Department of Chemistry and Center for Nucleic Acids Science and Technology, Carnegie Mellon University , 4400 Fifth Avenue, Pittsburgh, Pennsylvania 15213-3890, United States.
authors.forename: Marcel P
authors.initials: MP
authors.lastname: Bruchez
authors.affiliation: Department of Chemistry and Center for Nucleic Acids Science and Technology, Carnegie Mellon University , 4400 Fifth Avenue, Pittsburgh, Pennsylvania 15213-3890, United States.
authors.forename: Bruce A
authors.initials: BA
authors.lastname: Armitage
mesh_terms: D020121:5' Untranslated Regions / Q000187:drug effects; D020816:Amino Acid Motifs; D000818:Animals; D015195:Drug Design; D054856:G-Quadruplexes / Q000187:drug effects*; D020558:GTP Phosphohydrolases / Q000235:genetics; D017930:Genes, Reporter / Q000187:drug effects; D005998:Glycine / Q000031:analogs & derivatives / Q000737:chemistry; D006801:Humans; D007700:Kinetics; D008565:Membrane Proteins / Q000235:genetics; D009690:Nucleic Acid Conformation; D009691:Nucleic Acid Denaturation; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*; D014176:Protein Biosynthesis / Q000187:drug effects*; D011500:Protein Synthesis Inhibitors / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D020871:RNA Stability / Q000187:drug effects; D012333:RNA, Messenger / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism; D011817:Rabbits; D012156:Reticulocytes / Q000201:enzymology / Q000378:metabolism
chemical_list: D020121:5' Untranslated Regions; D008565:Membrane Proteins; D016376:Oligonucleotides, Antisense; D011500:Protein Synthesis Inhibitors; D012333:RNA, Messenger; C028791:N-(2-aminoethyl)glycine; D020558:GTP Phosphohydrolases; C579846:NRAS protein, human; D005998:Glycine