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US 12,753,426 B2Granted
High throughput assays for detecting translation readthrough inducers
AssigneeThe Trustees of the University of Pennsylvania
InventorsBarry S. Cooperman, Mikel D. Ghelfi
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Abstract
Disclosed are systems and methods for high throughput screening to detect translation readthrough induced drugs (TRID). The systems use highly purified, eukaryotic cell-free protein synthesis systems that distinguish TRIDs acting directly on the protein synthesis machinery from those that act indirectly.
Claims20
A system comprising a substrate and a plurality of regions on the substrate, each region comprising a cell-free ribosome-dependent protein synthesis reporter complex, wherein the cell-free ribosome-dependent protein synthesis reporter complex comprises:
- a) transfer-ribonucleic acid linked to a peptide (peptidyl-tRNA), wherein one or more amino acids of the peptide are labeled with a first fluorophore or a chromophore,
- b) a transfer-ribonucleic acid;
- c) a modified ribonucleic acid (RNA) molecule comprising an internal ribosome entry site (IRES) linked to a coding region encoding the peptide of the peptidyl-tRNA and a termination codon (Stop-IRES mRNA);
- and
- d) a ribosome,
- wherein the transfer-ribonucleic acid or the ribosome is linked to a second fluorophore or a second chromophore to form a labeled-tRNA or a labeled ribosome, respectively.
3 more elements
Dependencies are inferred from the claim text.