NOVEL IN VITRO EXPRESSION METHODS FOR PROTEOME ANALYSIS

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Proteonomic analysis is an important goal for the post-genome era. While a complete list of genes from different organisms is rapidly being compiled, the function of most of these genes remains undetermined. This knowledge is crucial in areas ranging from drug discovery to understanding the basic mechanisms of cellular regulation. We will evaluate a new approach for proteomic analysis based on recent progress in the area of in vitro expression and isolation of proteins. Until now, it has not been possible to isolate proteins which are produced in a cell- free extract in an unaltered form. However, analysis of functional libraries of in vitro expressed proteins (LIVE-PRO) derived from mRNA libraries of tissues and individual cells offers many advantages. The incorporation of proprietary photocleavable biotins through enzymatically and chemically aminoacylated tRNAs solves this problem by facilitating the rapid isolation of an in vitro expressed protein library, even for proteins with low copy number. Importantly, this approach facilitates the downstream analysis and mapping of the proteome through a variety of techniques including 2D-gels, mass spectrometry, affinity binding and biochemical assays. Drug discovery will be facilitated by observing differential effects on in vitro expressed proteins and by downstream functional analysis of drug-protein interactions. PROPOSED COMMERCIAL APPLICATIONS: The development of new methods for the isolation, detection and analysis of in vitro expressed proteins will provide a new path for mapping and characterizing an organisms proteome. Such knowledge is important for both understanding basic cellular mechanisms and the development of new drugs. Products include kits and systems for high throughput screening of in vitro expressed libraries of proteins and new methodology for drug discovery. See attached support letter from Promega.