Dna construct for assessing thymic function activity and...

C - Chemistry – Metallurgy – 12 – N

Patent

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C12N 15/10 (2006.01) C12N 15/63 (2006.01) C12N 15/64 (2006.01) C12N 15/85 (2006.01) C12Q 1/68 (2006.01) C12N 5/06 (2006.01)

Patent

CA 2467858

The present invention relates to a DNA construct for in vivo expression in an excision DNA circle created by DNA recombination machinery in T cells from a non-human mammal which comprises two recombination signal sequences (RSS) consensus sequences flanking a promoter, an enhancer and a reporter gene, wherein the excision DNA circle is diluted out after cellular division and the excision DNA circle is detected by expression of the reporter gene and the detection is indicative of thymic function activity of the mammal. The present invention also relates to a T cell transiently transfected with the DNA construct of the present invention, the cell expressing quantifiable levels of reporter gene for green fluorescent protein (GFP) for determining enhancing/decreasing thymic exportation, a mammal comprising the DNA construct of the present invention and methods thereof.

La présente invention concerne un ADN recombiné permettant d'exprimer, in vivo, un cercle d'excision d'ADN, créé par technique de recombinaison d'ADN sur des lymphocytes T prélevés sur un mammifère non humain, comprenant deux séquences consensus de séquences spécifiques de recombinaison (RSS) flanquant un promoteur, un amplificateur et un gène rapporteur. Le cercle d'excision d'ADN est dilué après la division cellulaire et détecté par expression du gène rapporteur, la détection permettant de mesurer l'activité de la fonction thymique du mammifère. La présente invention concerne également un lymphocyte T transfecté de façon transitoire avec l'ADN recombiné de la présente invention, la cellule exprimant des niveaux quantifiables du gène rapporteur pour une protéine verte fluorescente (GFP) permettant de mesurer l'augmentation ou la diminution de l'exportation thymique. L'invention concerne également l'ADN recombiné de la présente invention et des applications thérapeutiques associées.

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