C - Chemistry – Metallurgy – 12 – Q
Patent
C - Chemistry, Metallurgy
12
Q
C12Q 1/68 (2006.01) C12N 9/10 (2006.01) C12N 9/12 (2006.01) C12N 9/14 (2006.01) C12N 15/81 (2006.01)
Patent
CA 2349827
This invention provides methods for the discovery of molecules that target an essential aspect of eukaryotic gene expression - the formation of the mRNA 5' cap m7GpppN. An underlying principle of this invention is the use of different strains of a test organism that differ only in the composition or source of the essential cap-forming enzymes. The invention provides isogenic yeast strains that derive all their capping activities from fungal sources versus mammalian sources. These strains form the basis of a differential growth inhibition assay to identify molecules that specifically target the fungal capping apparatus. This invention also provides a method to screen in vitro for molecules that inhibit fungal RNA triphosphatase, an essential enzyme that catalyzes the first of three steps in cap synthesis.
L'invention concerne des procédés relatifs à la découverte de molécules qui ciblent un aspect essentiel de l'expression de gène eucaryote - à savoir la formation de coiffe m7GpppN d'extrémité 5' d'ARNm. Aux fins de l'invention, un principe sous-jacent est l'utilisation de diverses souches d'un organisme test qui diffère seulement dans la composition ou la source des enzymes essentielles pour la formation de coiffe. L'invention concerne en outre des souches de levure isogéniques qui dérivent toutes leurs activités de formation de coiffe de sources fongiques par opposition à des sources mammaliennes. Ces souches constituent la base d'un essai d'inhibition de croissance de type différentiel visant à identifier des molécules ciblant spécifiquement le dispositif de formation de coiffe fongique. L'invention concerne par ailleurs un procédé de criblage in vitro pour des molécules qui inhibent l'ARN triphosphatase fongique, une enzyme essentielle qui catalyse la première des trois étapes dans la synthèse de coiffe.
Borden Ladner Gervais Llp
Sloan-Kettering Institute For Cancer Research
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