Cell-based drug screens for regulators of gene expression

Details Cell-based drug screens for regulators of gene expression Cell-based drug screens for regulators of gene expression

C - Chemistry, Metallurgy

12

N

C12N 5/10 (2006.01) C12N 15/10 (2006.01) C12N 15/63 (2006.01) C12N 15/64 (2006.01) C12N 15/85 (2006.01) C12Q 1/66 (2006.01) C12Q 1/68 (2006.01) G01N 33/50 (2006.01)

Patent

CA 2265251

The invention provides methods and compositions for screening for pharmacological agents which regulate gene expression in mammals. An exemplary assay involves (a) contacting a mammalian cell comprising a knock-in mutant of a targeted native allele encoding a reporter of gene expression, as shown in the figure, wherein the expression of the reporter is under the control of the gene expression regulatory sequences of the native allele, with a candidate agent under conditions whereby but for the presence of the agent, the reporter is expressed at a first expression level; and (b) measuring the expression of the reporter to obtain a second expression level, wherein a difference between the first and second expressions levels indicates that the candidate agent modulates gene expression.

L'invention concerne des procédés et des compositions destinés au criblage d'agents pharmacologiques régulant l'expression génétique chez les mammifères. Un dosage effectué à titre d'exemple consiste à: (a) mettre en contact une cellule mammalienne comprenant un mutant par choc d'un allèle endogène ciblé codant pour un marqueur d'expression génétique, comme indiqué dans le schéma 1, l'expression du marqueur étant régulée par les séquences de régulation d'expression génétique de l'allèle endogène, avec un agent candidat dans des conditions telles que, sans la présence de l'agent, le marqueur est exprimé à un premier niveau d'expression; et (b) mesurer l'expression du marqueur afin d'obtenir un deuxième niveau d'expression, dans lequel une différence entre le premier et le deuxième niveau d'expression indique que l'agent candidat module l'expression génétique.

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