At the present time it is not possible to rapidly and inexpensively obtain a vertebrate knockout in a gene of choice, or to readily perform whole genome forward genetic screening in a vertebrate. The goal of this Phase II SBIR application is to create the research tools necessary to perform these functional genomics experiments in a vertebrate by developing a zebrafish repository containing a tagged insertional mutation in every identifiable gene. The proposed research will focus on creating, storing, and managing a zebrafish sperm library, termed the Zenemark Library, containing retrovirally-mediated insertional mutations in each lene in the zebrafish genome. Each of these insertional mutations will be characterized, stored as multiple =liquots of frozen sperm, and entered in a databank using identifiers of flanking genomic sequence, zebrafish gene locus, and mammalian gene homologues. A collection of 1,000,000 independent inserts will create, on average, a tagged genetic disruption once every 1800 base pairs, essentially a mutation in every zebrafish gene. The database, and individual mutant strains derived from the library will be available to both commercial and academic researchers. Just as this resource will allow basic researchers to more readily study the genetics of biological processes in vertebrates, it will allow pharmaceutical researchers to apply whole genome forward genetics to the problems of drug and drug target discovery. In particular, the creation of the ZeneMark Library will allow pharmaceutical researchers to screen collections of mutant fish, such as fish with mutations in all druggable genes, to 1 ) identify novel drug targets, 2) identify zebrafish models of human disease that may be used directly for high-throughput in vivo drug discovery, and 3) identify the site(s) of action of known drugs, toxins, or other chemicals. This forward genetic screening will serve as the primary basis for commercialization of the library, and Znomics is already in the process of seeking both corporate partners as well as venture capital to fund Phase III of this program. ? ?
