A Lack of premature termination codon read-through efficacy of PTC124 (Ataluren) in a diverse array of reporter assays

Stuart P. McElroy, Toshifumi Nomura, Leah S. Torrie, Emma Warbrick, Ulrike Gartner, Gavin Wood, W.H. Irwin McLean

    Research output: Contribution to journalArticle

    78 Citations (Scopus)

    Abstract

    The drug molecule PTC124 (Ataluren) has been described as a read-through agent, capable of suppressing premature termination codons (PTCs) and restoring functional protein production from genes disrupted by nonsense mutations. Following the discovery of PTC124 there was some controversy regarding its mechanism of action with two reports attributing its activity to an off-target effect on the Firefly luciferase (FLuc) reporter used in the development of the molecule. Despite questions remaining as to its mechanism of action, development of PTC124 continued into the clinic and it is being actively pursued as a potential nonsense mutation therapy. To thoroughly test the ability of PTC124 to read through nonsense mutations, we conducted a detailed assessment comparing the efficacy of PTC124 with the classical aminoglycoside antibiotic read-through agent geneticin (G418) across a diverse range of in vitro reporter assays. We can confirm the off-target FLuc activity of PTC124 but found that, while G418 exhibits varying activity in every read-through assay, there is no evidence of activity for PTC124.
    Original languageEnglish
    Article numbere1001593
    JournalPLoS Biology
    Volume11
    Issue number6
    DOIs
    Publication statusPublished - 2013

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    nonsense mutation
    stop codon
    Nonsense Codon
    Assays
    luciferase
    mechanism of action
    assays
    geneticin
    aminoglycoside antibiotics
    Firefly Luciferases
    drugs
    therapeutics
    Molecules
    ataluren
    Aminoglycosides
    genes
    proteins
    testing
    Genes
    Anti-Bacterial Agents

    Cite this

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    abstract = "The drug molecule PTC124 (Ataluren) has been described as a read-through agent, capable of suppressing premature termination codons (PTCs) and restoring functional protein production from genes disrupted by nonsense mutations. Following the discovery of PTC124 there was some controversy regarding its mechanism of action with two reports attributing its activity to an off-target effect on the Firefly luciferase (FLuc) reporter used in the development of the molecule. Despite questions remaining as to its mechanism of action, development of PTC124 continued into the clinic and it is being actively pursued as a potential nonsense mutation therapy. To thoroughly test the ability of PTC124 to read through nonsense mutations, we conducted a detailed assessment comparing the efficacy of PTC124 with the classical aminoglycoside antibiotic read-through agent geneticin (G418) across a diverse range of in vitro reporter assays. We can confirm the off-target FLuc activity of PTC124 but found that, while G418 exhibits varying activity in every read-through assay, there is no evidence of activity for PTC124.",
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    A Lack of premature termination codon read-through efficacy of PTC124 (Ataluren) in a diverse array of reporter assays. / McElroy, Stuart P.; Nomura, Toshifumi; Torrie, Leah S.; Warbrick, Emma; Gartner, Ulrike; Wood, Gavin; McLean, W.H. Irwin.

    In: PLoS Biology, Vol. 11, No. 6, e1001593, 2013.

    Research output: Contribution to journalArticle

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