SBIR-STTR Award

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 this transforms 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 research is 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 of large 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 new industrial 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 large quantities 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 $.05 each. 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

Keywords:
Transgenic, Pharmaceuticals, Poultry, Genes, Proteins, Bioreactor, Transposons, Testes

The objective is completion of the project, initiated in Phase I, to generate transgenic chickens capable of expressing and secreting specified proteins into the whites of eggs, allowing economical large scale production. In the Phase I effort, seventeen birds were shown to have generated transgenic sperm as demanded by the criteria for success specified in the research protocols. This satisfies our Phase I objective. It is intended that the Phase I results will be extended in Phase II to generate avians capable of producing large quantities of pharmaceutical proteins (vaccines, antibodies, antidotes, etc.) in their eggs to allow efficient and economical production for commercial purposes as well as Public Health and Defense Department needs. The Company has already received contacts from industrial concerns interested in utilizing this technology for gaining access to otherwise unobtainable markets. In one instance the first year market for their first product would approximate $160M. Avian transgenic technology has the potential to provide vast quantities of protein products used in vaccines, biocides, antidotes and many as yet undiscovered applications owing to its enhanced and accessible scale of protein production coupled to its vastly lower cost for production. The modular character of the transgenic vector allows alternative protein expression systems to be introduced within the vector's genetic framework, to provide a versatile technique for altering production objectives to accommodate changing customer requirements. People are increasingly concerned about the current world which witnesses rapid dissemination of new diseases by commercial transportation and the potential of bioterroism by sophisticated international criminals. Some mechanisms are required to provide large amounts of effective counteracting drugs. One immediate need for such agents is their use by our armed forces which must travel to remote regions of the world where exotic diseases flourish. Not only are large numbers of troops in need of such protection, it is also in our country's interest to prevent rapidly spreading plagues from dispersing throughout the world. We believe that not only does transgenic poultry technology provide this production potential, it also has the ability to adapt rapidly to changing production requirements.

Keywords:
transgenic, chickens, transposon, biomanufacturing,egg white, biopharmaceuticals