1. Chemical-Induced Read-Through at Premature Termination Codons Determined by a Rapid Dual-Fluorescence System Based on S. cerevisiae.
- Author
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Altamura, Emiliano, Borgatti, Monica, Finotti, Alessia, Gasparello, Jessica, Gambari, Roberto, Spinelli, Mariangela, Castaldo, Rosa, and Altamura, Nicola
- Subjects
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NONSENSE mutation , *STOP codons , *MESSENGER RNA , *SACCHAROMYCES cerevisiae , *QUANTITATIVE research - Abstract
Nonsense mutations generate in-frame stop codons in mRNA leading to a premature arrest of translation. Functional consequences of premature termination codons (PTCs) include the synthesis of truncated proteins with loss of protein function causing severe inherited or acquired diseases. A therapeutic approach has been recently developed that is based on the use of chemical agents with the ability to suppress PTCs (read-through) restoring the synthesis of a functional full-length protein. Research interest for compounds able to induce read-through requires an efficient high throughput large scale screening system. We present a rapid, sensitive and quantitative method based on a dual-fluorescence reporter expressed in the yeast Saccharomyces cerevisiae to monitor and quantitate read-through at PTCs. We have shown that our novel system works equally well in detecting read-through at all three PTCs UGA, UAG and UAA. [ABSTRACT FROM AUTHOR]
- Published
- 2016
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