Gene disruption methodologies for drug target discovery

C - Chemistry – Metallurgy – 12 – N

Patent

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C12N 1/15 (2006.01) A61K 39/00 (2006.01) C07K 14/40 (2006.01) C07K 16/14 (2006.01) C12N 15/10 (2006.01) C12N 15/65 (2006.01) C12N 15/80 (2006.01) C12N 15/90 (2006.01) C12P 21/00 (2006.01) G01N 33/50 (2006.01)

Patent

CA 2398861

The present invention provides methods and compositions that enable the experimental determination as to whether any gene in the genome of a diploid pathogenic organism is essential, and whether it is required for virulence or pathogenicity. The methods involve the construction of genetic mutants in which one allele of a specific gene is inactivated while the other allele of the gene is placed under conditional expression. The identification of essential genes and those genes critical to the development of virulent infections, provides a basis for the development of screens for new drugs against such pathogenic organisms. The present invention further provides <i>Candida albicans</i> genes that are demonstrated to be essential and are potential targets for drug screening. The nucleotide sequence of the target genes can be used for various drug discovery purposes, such as expression of the recombinant protein, hybridization assay and construction of nucleic acid arrays. The uses of proteins encoded by the essential genes, and genetically engineered cells comprising modified alleles of essential genes in various screening methods are also encompassed by the invention.

L'invention concerne des procédés et compositions permettant de déterminer de manière expérimentale si un quelconque gène du génome d'un organisme pathogène diploïde est essentiel et s'il est nécessaire à la virulence ou au pouvoir pathogène. Ces procédés consistent à construire des mutants génétiques dans lesquels un allèle d'un gène spécifique est inactivé tandis que l'autre allèle du gène est placé dans des conditions d'éventuelle expression. L'identification de gènes essentiels et de gènes critiques quant au développement d'infections virulentes constitue une base de développement du criblage de nouveaux médicaments dirigés contre ces organismes pathogènes. L'invention concerne encore des gènes <i>Candida albicans</i> qui se sont révélés être essentiels dans le criblage de médicaments et constituent des cibles potentielles à cette fin. On peut utiliser la séquence nucléotidique de ces gènes cibles à des fins de découverte de médicaments, telle que l'expression de la protéine de recombinaison, le dosage d'hybridation et la construction d'ensembles d'acides nucléiques. L'invention concerne enfin l'utilisation de protéines, codées par les gènes essentiels, et de cellules modifiées génétiquement et comprenant des allèles modifiés de gènes essentiels, dans divers procédés de criblage.

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