Transcriptional activators with graded transactivation...

C - Chemistry – Metallurgy – 12 – N

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C12N 15/62 (2006.01) A61K 48/00 (2006.01) C07K 14/035 (2006.01) C07K 19/00 (2006.01) C12N 5/10 (2006.01) C12N 15/38 (2006.01) C12N 15/63 (2006.01) C12N 15/67 (2006.01) C12N 15/85 (2006.01)

Patent

CA 2294598

Transcriptional activators which differ in their activation potential by more than 3 orders of magnitude are provided. The transactivators are fusions between a DNA binding protein (e.g., a Tet repressor) and minimal transcriptional activation domains derived from Herpes simplex virus protein 16 (VP16). Substitution mutations at amino acid position 442 within the minimal VP16 domain provide transactivators with differing transactivation ability. Moreover, chimeric activation domains comprising both wild type and mutant minimal VP16 domains provide additional variants with differing transactivation ability. Various aspects of the invention pertain to nucleic acid molecules, vectors, host cells, fusion proteins, transgenic and homologous recombinant organisms and methods of regulating gene transcription.

La présente invention concerne des activateurs de transcription ayant une différence de potentiel d'activation dépassant les trois ordres d'amplitude. Les transactivateurs sont le résultat de fusions entre une protéine de liaison d'ADN (par exemple un répresseur de Tet) et des domaines d'activation transcriptionnels minimums obtenus à partir de la protéine 16 du virus de l'herpès simplex (VP16). Les mutations par substitution au niveau de la position 442 de l'acide aminé dans le domaine minimum VP16 fournissent des transactivateurs ayant des capacités de transactivation différentes. De plus, les domaines d'activation chimériques comprenant des domaines VP16 minimums à la fois de type sauvage et mutant fournissent des variants supplémentaires avec des capacités de transactivation différentes. Différents aspects de l'invention concernent des molécules d'acide nucléique, des vecteurs, des cellules hôtes, des protéines de fusion, des organismes de recombinaison homologues et transgéniques ainsi que des méthodes de régulation de la transcription génique.

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