A reporter system for translational readthrough of stop codons in human cells

Agents to induce readthrough of premature termination codons (PTCs) are useful research tools and potential therapeutics. Reporters used to detect PTC readthrough are gene-specific and thus are not suited to for general assessment of readthrough activity or in cases where PTC-inactivated genes are unknown. Here we describe a GFP-based reporter construct pMHG-W57∗ which is capable of detecting dose-dependent drug-induced PTC readthrough both by fluorescence microscopy and flow cytometry. pMHG-W57∗ may be used as a general indicator of PTC readthrough in living cells and obviates the need for gene-specific recoding sequences in reporter constructs.

▸ Current reporter systems for translational readthrough are gene-specific and cannot be used in living cells. ▸ We developed a readthrough reporter based on green fluorescent protein. ▸ The reporter is gene-independent and can be used to sort living cells based on fluorescence.