Cotranslational Protein Expression System for High-throughput Screening

Reporter gene-based assays are used extensively in high-throughput screening (HTS) to identify chemical modulators of cellular pathways for drug discovery and development. However, such screening frequently results in a large number of false “hits” due to interactions of screened compounds with reporter proteins, producing confounding results. Thus, validation of results using these assays often involves significant time and expense.

The inventors have developed an assay system that significantly improves detection of target-relevant active compounds by discriminating between signals arising from the target activity and those caused by reporter bias. This system utilizes simultaneous detection (also known as “coincidence detection”) of non-homologous reporter proteins with dissimilar properties, such as differing catalysis, light emission, or fluorescence characteristics; simultaneous observation of signals from these reporters indicates a high probability that it is a true target response. The reporters are cotranslationally expressed from a single RNA transcript, which ensures stable stoichiometry of the expressed proteins.

Potential Commercial Applications: Competitive Advantages:
High-throughput screening of chemical libraries in a single assay platform for commercial or research use.   This method will significantly enhance the ability to identify and prioritize active compounds from reporter gene-based assays.

James Inglese (NCATS)
Chih-Chien (Ken) Cheng (NCATS)
Samuel Hasson (NCATS)

Intellectual Property:
PCT Application No. PCT/US2013/032184

Collaboration Opportunity:

The National Center for Advancing Translational Sciences (NCATS) is seeking statements of capability or interest from parties interested in collaborative research to further develop, evaluate or commercialize Cotranslational Protein Expression System for High-throughput Screening. For collaboration opportunities, please contact NCATS Technology Development Coordinator at

Licensing Contact:
Tara Kirby , Ph.D.
NIH Office of Technology Transfer
Phone: 301-435-4426

OTT Reference No: E-300-2012/0

Updated: Aug-06-2013