The EST (expressed sequence tag) paradigm to randomly sequence short stretches of expressed regions of genes as a means to identify full-length cDNAs, has been challenged by the lack of technological innovations to pick up where this technology left off. The industry is now guttered with millions of ESTs but is unable to extend this discovery process to the identification and characterization of full-length genes. OriGene Technologies, Inc. (OTI) has developed two complementary technologies which allow high-throughput full-length cDNA cloning. In this fast-track Phase I - Phase II application, OTI proposes a gene discovery program that targets human prostate cancer. In the Phase I component, the applicants propose to generate high-quality source material which will support large-scale, full-length cDNA isolation at the Phase II stage.
Three specific aims are proposed: (1) generating cDNA libraries from normal prostate tissue and prostatic adenocarcinomas, (2) arraying the cDNA clones from each library onto primary and secondary multi-level plates, and (3) performing stringent quality controls on these library. Prostate carcinoma is the most frequently diagnosed form of human cancer in the United States. The applicants assert that many genes which function in prostate carcinogenesis could serve as useful diagnostic tools and/or as targets for therapeutic intervention. In the Phase II component, source materials developed in the Phase I component will be used to isolate two hundred full-length cDNA clones, selected on the basis of their prostate-specific or prostate cancer-specific expression patterns. The applicants propose: (1) developing approaches to select important ESTs that potentially play a role in prostate growth and carcinogenesis, (2) isolating their corresponding full-length cDNA clones from the arrayed cDNA library panels, and (3) determining the 5' DNA sequence of each and confirming its authenticity. This discovery process is likely to extend beyond the work proposed in this application.
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