A cytotoxicity-based genetic selection system (toxsel)

C - Chemistry – Metallurgy – 12 – Q

Patent

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Details

C12Q 1/02 (2006.01) C12N 15/10 (2006.01) C12Q 1/68 (2006.01) G01N 33/50 (2006.01)

Patent

CA 2222941

The invention provides a cytotoxicity-based genetic selection (TOXSEL) method and related testing kit for the identification and positive selection of molecules or mutations that are capable of disrupting a specific protein- protein interaction. The invention enables positive selection of molecules disruptive of specific protein-protein interactions by virtue of the presence in the TOXSEL system of a toxin reporter gene. A disrupted protein-protein interaction precludes expression of the toxin reporter gene and, consequently, allows survival of the host cell. TOXSEL technology enables large-scale screening for drugs or small molecules capable of disrupting specific protein- protein interactions critical in processes such as cellular signalling, carcinogenesis, etc. The positive selection of molecules or mutations capable of interfering with specific protein-protein interactions permits the identification of amino acid residues or motifs critical in the protein- protein interactions and permits the design of drugs for therapeutic or diagnostic applications.

L'invention porte sur une méthode de sélection génétique basée sur la cytotoxicité (TOXSEL) et son matériel d'expérimentation destinés à l'identification et la sélection positive de molécules ou de mutations à même d'interrompre une interaction inter-protéique spécifique. L'invention permet la sélection positive de molécules qui interrompent les interactions inter-protéiques spécifiques grâce à la présence dans le système TOXSEL d'un gène reporter de toxine. Une interaction interprotéique interrompue empêche l'expression du gène reporter de toxine et, par conséquent, permet d'assurer la survie de la cellule hôte. La technologie TOXSEL permet le criblage à grande échelle de médicaments ou de petites molécules à même d'interrompre des interactions inter-protéiques spécifiques indispensables aux processus de signalisation cellulaire, de carcinogenèse, etc. La sélection positive de molécules ou de mutations à même de faire obstacle aux interactions inter-protéiques spécifiques permet d'identifier des restes ou des motifs structuraux d'acides aminés et de concevoir des médicaments à des fins thérapeutiques ou diagnostiques.

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