Methods of assaying for compounds that inhibit premature translation termination and nonsense-mediated RNA decay
OwnerHolger Beckmann, Marc Learned +2inventors
InventorsHolger Beckmann, Marc Learned, Stuart Peltz, Kevin Czaplinski
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Abstract
The present application provides methods of assaying for compounds that inhibit premature translation termination and nonsense mediated RNA decay in cells.
Claims32
A method of in vitro screening for compounds that modulate premature translation termination and nonsense-mediated mRNA decay, the method comprising the steps of:
- (i) incubating a translation assay, the assay comprising an in vitro translation cellular extract;
- a nucleic acid encoding a polypeptide, wherein the coding sequence for the polypeptide comprises a premature stop codon;
- and a candidate modulator compound;
- and
- (ii) detecting the polypeptide translated from the nucleic acid.
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The method of in vivo screening for compounds that modulate premature translation termination and nonsense-mediated mRNA decay, the method comprising the steps of:
- (i) expressing in a cell a nucleic acid encoding a polypeptide, wherein the coding sequence for the polypeptide comprises a premature stop codon;
- (ii) contacting the cell with a candidate modulator compound;
- and
- (iii) detecting either the polypeptide translated from the nucleic acid or RNA transcribed from the nucleic acid.
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30-34. (canceled)
A kit for screening for compounds that modulate translation termination and nonsense-mediated mRNA decay, the kit comprising a nucleic acid encoding a polypeptide, wherein the polypeptide coding sequence comprises a premature stop codon;
- instructions for practicing a method of screening for compounds that inhibit translation termination at premature stop codons and nonsense mediated RNA decay;
- and a control compound that inhibits nonsense-mediated RNA decay.
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Dependencies are inferred from the claim text.