Patent ID: 7927791
Filing Date: 2011-04-19
Classification: B82Y,G01N

Abstract:
1. A method of identifying a small molecule that modulates premature translation termination or nonsense-mediated mRNA decay, said method comprising: (a) contacting a region of human 28S rRNA with a library of small molecules under conditions that permit direct binding of the region of human 28S rRNA to a member of the library of small molecules and the formation of a region of human 28S rRNA:small molecule complex; (b) detecting the formation of a region of human 28S rRNA:small molecule comple, wherein a small molecule that binds to the region of human 28S rRNA is identified if a region of human 28S rRNA:small molecule complex is detected; (c) contacting the small molecule identified as binding to a region of human 28S rRNA with a cell-free extract and a nucleic acid sequence comprising a regulatory element operably linked to a reporter gene coding region, wherein the reporter gene coding region comprises a premature stop codon, and wherein the cell-free extract is isolated from cells that have been incubated on ice for at least 12 hours; and (d) detecting the protein expressed from the reporter gene coding region, wherein a small molecule that modulates premature translation termination or nonsense-mediated mRNA decay is identified if the protein expressed from the reporter gene coding, region in the presence of the small molecule is altered relative to the protein expressed from the reporter gene coding region in the absence of the small molecule or the presence of a negative control.