System and method for regulation of gene expression

C - Chemistry – Metallurgy – 12 – N

Patent

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Details

C12N 15/85 (2006.01) C12N 5/10 (2006.01) C12N 15/63 (2006.01) C12N 15/67 (2006.01) C12Q 1/68 (2006.01)

Patent

CA 2316189

A control system for genetic expression includes a first, reporter polynucleotide molecule including a reporter sequence encoding a protein to be expressed operably linked to a heptameric tet operator sequence and at least two promoters and a second, regulator polynucleotide molecule including a sequence coding for a tet repressor protein and the KRAB protein. The reporter polynucleotide includes one promoter on each side of the tet operator sequence. In one embodiment a TK promoter is located upstream of the operator sequence and a CMV promoter is located downstream thereof. The desired nucleic acid sequence of the reporter molecule is positioned downstream of the CMV promoter. A method to regulate the expression of a protein using the system of the invention is provided. The invention also provides a kit including both the polynucleotides described above.

L'invention concerne un système de régulation de l'expression génique qui comprend une première molécule polynucléotidique marqueur comprenant une séquence marqueur codant une protéine devant être exprimée liée de manière opérationnelle à une séquence heptamère opérateur tet et à au moins deux promoteurs et une seconde molécule polynucléotidique régulatrice comprenant une séquence codant pour un répresseur tet et la protéine KRAB. Le polynucléotide marqueur comprend un promoteur sur chaque côté de la séquence opérateur tet. Selon une variante, un promoteur TK est placé en amont de la séquence opérateur et un promoteur CMV est placé en aval de celle-ci. La séquence d'acides nucléiques souhaitée de la molécule marqueur est placée en aval du promoteur CMV. Un procédé de régulation de l'expression d'une protéine au moyen du système fait aussi l'objet de cette invention. L'invention concerne en outre un ensemble comprenant les deux polynucléotides décrits.

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