BACKGROUND:  Understanding protein function and interaction with other cellular components on a genome-wide scale is one of the central goals of systems biology. Despite advances in high-throughput technologies, library-against-library screening with proteome-wide coverage remains a daunting task due to the huge demand of time and resources.

Previously, UCLA researchers have developed a new pooling-deconvolution method to pool "prey" or "bait" proteins. The method assigned 2n bait (BD) strains into n pairs of pools and screened the bait pools against prey (AD) arrays. However, bait pooling did not provide as large a benefit to research programs.

INNOVATION:  UCLA investigators are applying the same pooling-deconvolution principle to pool prey (AD) strains, instead of bait (BD) strains. Now, individual baits can be screened against a prey-based pool. Prey-based pools are advantageous over bait-based pools because prey pool arrays can be maintained indefinitely and reused for new screens. Due to this new prey-based pool approach, UCLA researchers have constructed a novel S. cerevisiae genome, two-hybrid smart pool array (SPA) system that increases yeast two-hybrid screening efficiency by an order of magnitude.

POTENTIAL APPLICATIONS 

• Protein interactome mapping
• Large-scale drug or drug target screening
• Screen small molecules that disrupt protein-protein interactions

ADVANTAGES

• Identifies protein interactions accurately with fewer experiments and less resources
• Combines advantages of both two-hybrid array screening and library screening approaches
• Prey-based strain pools can be maintained indefinitely and used repeatedly
• Increases screening efficiency by an order of magnitude and improves data quality
• Built-in redundancy removes false positive signals
• Pooling of arrays reduces array size and does not compromise the sensitivity of detecting reproducible hits

DEVELOPMENT-TO-DATE:  In addition to previous successful validations of the pooling-deconvolution method, researchers have constructed the first yeast two-hybrid SPAs of the S. cerevisiae genome. Five bait strains have been screened against SPAs with different pool sizes.

Related Papers (Selected) Jin F, Hazbun T, Michaud GA, Salcius M, Predki, Fields S, Huang J. A pooling-deconvolution strategy for biological network elucidation. Nature Methods. 3(3), 183-189 (2006). [more] Jin F, Avramova L, Huang J, Hazbun T. A yeast two-hybrid smart-pool-array system for protein-interaction mapping. Nature Methods 4(5), 405-407 (2007) [more]

Reference: UCLA Case No. 2006-089

PCT Application: US07/004313