In vivo screening of protein-protein interactions with protein-fragment complementation assays
US7855167B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Dec 5, 2003 |
| Grant date | Dec 21, 2010 |
| Priority date | — |
| Expiry date | Jan 13, 2026 |
Classification
- Technology area (CPC G)Physics
- CPC primaryG01N2800/52
- WIPO fieldMeasurement
- WIPO sectorInstruments
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.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.