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US 6,635,440 B1Granted
Method of isolation of RNA-binding compounds
AssigneeEuropean Molecular Biology Laboratory
InventorsMatthias W. Hentze, Efrosyni Paraskeva
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Abstract
The present invention provides a method of isolation of compounds that bind specifically to RNA or that alter the affinity of known compounds for RNA. The method utilizes changes in the levels of a fluorescent marker protein expressed in the same cell as a putative RNA-binding compound to detect those compounds of interest.
Claims11
A method for the identification of a compound that alters the affinity of interaction between an RNA-binding site and an RNA-binding compound, said method comprising the steps of:
- a) expressing in a host cell
- i) a nucleic acid comprising a coding region for the RNA-binding compound,
- ii) a nucleic acid comprising a transcribed sequence comprising a coding region for a fluorescent marker protein, wherein the nucleic acid comprising the coding region for the fluorescent marker protein includes a binding site for the RNA-binding compound within a transcript transcribed from the transcribed sequence, wherein binding of the RNA-binding compound to the RNA-binding site represses translation of the transcribed sequence,
- b) contacting said host cell with a compound to be screened for altering the affinity of interaction between the RNA-binding site and the RNA-binding compound, and
- c) measuring the level of fluorescence in said host cells, wherein a compound that alters said level of fluorescence from the level that is seen in the absence of compound is indicative of a compound that alters the affinity of interaction between the RNA-binding site and the RNA-binding compound.
2 more elements
Dependencies are inferred from the claim text.