SBIR-STTR Award

RNA-protein interactions play critical roles in cellular regulatory processes and in the life cycles of many pathogenic viruses, including HIV, hepatitis C, and West Nile virus, and thus represent attractive targets for therapeutics. Much of our previous work has centered on identifying and characterizing the components of key interactions and developing cell-based reporters to accurately monitor RNA-protein interactions. Here we focus on developing targeted assays for inhibitor screening. The cell-based screens described incorporate a system of genetic reporters designed to recapitulate RNA-protein interactions in a native mammalian environment and to enable screens with high sensitivity and specificity. As proof-of-principle, reporters cell lines will be developed and used to identify inhibitors of the HIV Rev-Rev Response Element (RRE) interaction from combinatorial zinc finger libraries. The development of high-throughput genetic methods to identify specific inhibitors of RNA-protein interactions is expected to help in the discovery of drugs directed against novel therapeutic targets.

Thesaurus Terms:
RNA, inhibitor /antagonist, molecular genetics, protein protein interaction, technology /technique development complementary DNA, genetic transcription, genetic translation, protein structure, reporter gene flow cytometry, high throughput technology, northern blotting, polymerase chain reaction

RNA-protein interactions play critical roles in cellular regulatory processes and in the life cycles of many pathogenic viruses including HIV, hepatitis C, SARS, and West Nile virus, and thus represent attractive targets for therapeutics. We have developed novel cell-based assays to identify drugs that target the RNA-protein interactions. The assays incorporate a system of genetic reporters to accurately recapitulate RNA-protein interactions and enable screens with high sensitivity and specificity. In an initial test of the system, the HIV Rev-RRE interaction was successfully targeted with control inhibitors of the RNA-protein interaction. Experiments to optimize and convert the drug screening assays to high throughput format are currently in progress. The assays will be tested by screening a combinatorial library of RRE-binding peptides to be followed by a high-throughput chemical compound library screen. The development of high-throughput genetic methods to identify specific inhibitors of RNA-protein interactions is expected to help in the discovery of drugs directed against novel therapeutic targets.

Thesaurus Terms:
RNA binding protein, gene expression, high throughput technology, human immunodeficiency virus, inhibitor /antagonist, intermolecular interaction, method development, molecular genetics alkaline phosphatase, antiAIDS agent, binding site, corticosteroid receptor, enzyme activity, genetic library, genetic promoter element, genetic transcription, luciferin monooxygenase, peptide library, reporter gene, small molecule RNA interference, cell line, flow cytometry