Reporter gene system for use in cell-based assessment of...

Details Reporter gene system for use in cell-based assessment of... Reporter gene system for use in cell-based assessment of...

C - Chemistry, Metallurgy

12

N

C12N 15/51 (2006.01) C07H 21/04 (2006.01) C07K 14/18 (2006.01) C12N 15/66 (2006.01) C12N 15/86 (2006.01) C12Q 1/37 (2006.01) C12Q 1/42 (2006.01) C12Q 1/68 (2006.01) G01N 33/50 (2006.01) G01N 33/576 (2006.01) G03C 5/00 (2006.01) C12Q 1/70 (2006.01)

Patent

CA 2328494

A cell-based assay system in which the detection of the reporter gene activity, or secreted alkaline phosphatase (SEAP), is dependent upon the protease activity of the Hepatitis C virus NS3 gene product. This system can be used to assess the activity of candidate protease inhibitors in a mammalian cell-based assay system. The assay system is simpler than previously described assays due to the use of SEAP which allows the reporter gene activity to be quantified by measuring the amount of secreted gene product in the cell media by monitoring the conversion of luminescent or colorimetric alkaline phosphatase substrate.

La présente invention concerne un système d'essai précaractérisé dans lequel la détection de l'activité gène rapporteur, ou de la phosphatase alcaline sécrétée (SEAP), est fonction de l'activité protéase du produit du gène NS3 du virus de l'hépatite C. Ce système permet de vérifier l'activité de candidats inhibiteurs de protéase dans un système d'essai précaractérisé de mammifère. Ce système d'essai est plus simple que les essais déjà connus en ce qu'il met en oeuvre la SEAP, ce qui permet de faire une évaluation quantitative de l'activité gène rapporteur en mesurant la quantité de produit génique sécrétée dans le milieu cellulaire, et ce, par simple surveillance de la conversion du substrat à phosphatase alcaline luminescente ou colorimétrique.

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