Bioengineered Proteins for Chemical/Biological Defense, Protection, and Decontamination

This proposal encompasses a facile method for generating transgenic chickens. Traditionally, the approach to accomplish this has been to target the avian egg or embryo for gene insertion using either viral or 'standard' plasmids as vectors. In contrast,our proposed method introduces two new features, i.e. (1) novel transposon DNA vectors and (2) injection of DNA into the testes of immature males. As a result, the injected genes will integrate into the genome of some proportion of spermatogonia and thistransforms a significant percentage of the eventual mature spermatozoa for transmission by normal breeding to the next generation. This procedure is expected to produce much higher efficiencies n the production of true germline transgenic offspring.Enhancement of integration of the injected trangene construct is accomplished by transposon vectors, which forces integration into the genome of forming spermatogenic cells, as demonstrated in fish by the Co-Investigator. Thus, this STTR proposed researchis an extension of previous work, which is now applied to avians. This simple and efficient methodology should facilitate generating transgenic poultry which secrete large amounts of recombinant proteins into their egg whites, thus providing a method oflarge scale, low cost protein manufacturing.At the end of Phase I we anticipate having 1- 10 true germline transgenic chicks which, after breeding, should demonstrate not only a new method for transgenesis but also provide the groundwork for a newindustrial niche using birds as large scale bioreactors to produce the myriad of recombinant proteins needed in the near future. TransWestTech, Inc. plans to be one of the pioneers in commercializing this technology to allow the production of largequantities of low cost proteins by contract, Once this approach has been confirmed, it should be possible to generate millions of transgenic hens within 18 months, each of which could secrete up to a gram of protein into each egg, at a cost of about $.05each. A small number of contract poultry producers could therefore produce multi-kilogram amounts of protein- pharmaceuticals annually for an estimated finished price of less than $1/gram