Screen for novel protein inhibitors based upon engineered...

C - Chemistry – Metallurgy – 12 – Q

Patent

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C12Q 1/68 (2006.01) G01N 33/68 (2006.01)

Patent

CA 2526012

The present invention provides a method for identifying the presence of a protein inhibitor of a target protein in a sample, comprising the steps of a) contacting said sample with a cell, wherein said cell contains i) an inducible lethal overactivity mutation in a gene affecting the target protein; and ii) a mutation in a second gene, wherein the activity of the target protein is essential to the cell and the mutation in the second gene functionally compensates for any reduction in the activity of the target protein; b) inducing the lethal overactivity mutation; and subsequently C) assessing protein inhibition by comparing the degree of survival of the cell in the presence and the absence of said sample. Also provided are cells for use in said method.

La présente invention porte sur un procédé visant à identifier la présence d'un inhibiteur d'une protéine cible dans un échantillon, ce procédé consistant à: a) mettre en contact l'échantillon avec une cellule, cette cellule contenant : i) une mutation de suractivité létale inductible dans un gène affectant la protéine cible; et ii) une mutation dans un second gène, l'activité de la protéine cible étant essentielle à la cellule et la mutation du second gène compense fonctionnellement toute réduction de l'activité de la protéine cible; b) induire la mutation de suractivité létale et ensuite C) évaluer l'inhibition de la protéine en comparant le degré de survie de la cellule en présence et en l'absence de l'échantillon. L'invention porte également sur des cellules destinées à être utilisées dans ce procédé.

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