Methods and kits for discovery of rna-binding compounds

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/68 (2006.01) C12Q 1/70 (2006.01)

Patent

CA 2331825

A solution-based assay for identifying RNA binding compounds, based on competition. The assay comprises a reporter molecule carrying a fluorescent or chromogenic group that can form a one-to-one complex with an RNA target molecule carrying a second fluorescent or chromogenic group in such a way that the two groups are in sufficient proximity for fluorescence resonance energy transfer and/or quenching to take place. Addition of a compound-to-be-tested prevents formation of the complex and thereby increases the fluorescence of the RNA target and/or reporter molecules relative to the signal obtained in the absence of the test compound. Quantitative screening methods are also included.

La présente invention concerne un dosage utilisant une solution permettant d'identifier des composés de fixation d'ARN, mis en compétition. Ce dosage renferme une molécule porteuse transportant un groupe fluorescent ou chromogène pouvant former un complexe à correspondance un pour un avec une molécule cible d'ARN transportant un second groupe fluorescent ou chromogène, de manière à ce que ces deux groupes soient suffisamment proches l'un de l'autre pour permettre le transfert et/ou l'extinction d'énergie de résonance par fluorescence. L'addition d'un composé devant être testé empêche la formation du complexe, augmentant ainsi la fluorescence des molécules cibles et/ou porteuses d'ARN, en fonction du signal obtenu en l'absence du composé d'analyse. Par ailleurs, cette invention concerne des procédés de sélection quantitative.

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