Assays to monitor amyloid precursor protein processing

Details Assays to monitor amyloid precursor protein processing Assays to monitor amyloid precursor protein processing

C - Chemistry, Metallurgy

12

N

C12N 15/62 (2006.01) C07K 14/47 (2006.01) C12N 5/10 (2006.01) C12N 15/12 (2006.01) C12N 15/79 (2006.01) C12Q 1/02 (2006.01)

Patent

CA 2477002

The present invention provides DNA constructs, genetically engineered host cells, and methods for identifying inhibitors of amyloid precursor protein (APP) processing. The methods provide for the convenient identification, in a single assay, of inhibitors of .beta.-secretase and .gamma.-secretase as well as other forms of APP processing. The methods rely on fusion proteins of APP and transcription factors in which APP processing releases the transcription factors, allowing the transcription factors to activate transcription of a reporter gene. Inhibitors are identified as substances that block or diminish transcription factor release from the fusion protein, thereby causing a diminution of reporter gene readout.

La présente invention concerne des ADN hybrides, des cellules hôtes génétiquement modifiées, ainsi que des méthodes servant à identifier des inhibiteurs de la maturation moléculaire de la protéine précurseur amyloïde (APP). Ces méthodes permettent d'identifier facilement, dans un seul dosage, des inhibiteurs de la bêta-sécrétase et de la gamma-sécrétase, ainsi que d'autres formes de la maturation moléculaire de l'APP. Ces méthodes reposent sur des protéines hybrides de APP et sur des facteurs de transcription dans lesquels la maturation cellulaire de APP libère les facteurs de transcription, permettant aux facteurs de transcription d'activer la transcription d'un gène rapporteur. Ces méthodes permettent d'identifier des inhibiteurs comme des substances qui bloquent ou réduisent la libération des facteurs de transcription par la protéine hybride, entraînant ainsi une réduction de la lecture du gène rapporteur.

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