In vivo screening of protein-protein interactions with...

Details In vivo screening of protein-protein interactions with... In vivo screening of protein-protein interactions with...

2003-12-05

2010-12-21

Liu, Sue (Department: 1639)

Combinatorial chemistry technology: method, library, apparatus

Method specially adapted for identifying a library member

Direct analysis of a library member, per se, by a physical...

C506S012000, C435S005000, C435S006120, C435S069700, C536S023100, C536S023500

Reexamination Certificate

active

07855167

ABSTRACT:
The present invention describes rapid methods to screen for biomolecular interactions in vivo based on protein fragment complementation assays (PCA). We have demonstrated an in vivo library-versus-library screening strategy that has numerous applications in the identification of novel protein-protein interactions and in directed evolution. Also we demonstrate the detection of protein-protein interactions starting with defined (full-length) cDNAs, and the concomitant generation of functional assays that provide initial validation of the cDNA products as being biologically relevant. Also, we screened a large cDNA collection using automated PCA, combined with quantitative detection of protein-protein complexes. The invention enables bait-vs.-library, library-vs.-library and defined gene screening in any type of cell or cellular context, and using a wide range of reporters and detection methods. The invention allows for identifying and validating genes involved in any cellular process and also provide assays to study effects of potential drugs, or gene knockouts on specific pathways.

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