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What is the role of readthrough therapy for nonsense mutations?

Suppressing translation termination selectively at PTCs (also referred to as readthrough) is a potential treatment strategy for genetic diseases caused by nonsense mutations, including CF13. Readthrough agents suppress termination at in-frame PTCs by promoting the accommodation of near-cognate aminoacyl-tRNAs into the ribosomal acceptor (A) site.

What percentage of reporter protein is produced by a stop codon?

In a study using a cell-free system, readthrough constructs containing a stop codon with a Ψ at the first nucleotide generated approximately 74–100% of the reporter protein produced by a sense construct. In contrast, a non-Ψ stop codon produced only ∼1% of wild-type protein (64).

What is the ribosome readthrough score?

The Ribosome ReadThrough Score (RRTS) was defined as the density of ribosomes in the 3´-UTR between the NTC and the first in-frame 3′-termination codon divided this value by the density of ribosomes in the CDS for every annotated transcript (as previously described36).

Does pseudouridylation of the first nucleotide of a stop codon lead to PTC readthrough?

In support of this hypothesis, recent studies have shown that pseudouridylation of the first nucleotide of a stop codon leads to PTC readthrough. In a study using a cell-free system, readthrough constructs containing a stop codon with a Ψ at the first nucleotide generated approximately 74–100% of the reporter protein produced by a sense construct.


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