Open Competition 3 - Biotechnology

Engineering Systems for Site Specific Alteration of Mammalian Genomes

Sponsor: Rheogene LLC

• Project Performance Period: 1/1/2002 - 7/31/2005
• Total project (est.): $3,358,382.00
• Requested ATP funds: $2,000,000.00

Existing methods for inserting genes into or deleting genes from chromosomes within an intact cell nucleus are either inefficient or unable to target specific sites. In fact, only one cell in a million undergoes the desired alteration. Furthermore, it is impossible to delete genes from cellular components called mitochondria. New methods are needed so that disease-causing genetic mutations can be identified and possibly corrected, and drug candidates can be screened faster and more effectively. RHeogene plans to develop and demonstrate tools for altering genetic material in either cell nuclei or mitochondra with at least 1,000 times the efficiency and specificity of existing methods. The three-year project will exploit the activities of naturally occurring enzymes. The company will identify enzymes called recombinases and integrases that have the capability to alter genes, and then identify the specific regions within the enzymes that perform these activities. Libraries of mutagenized forms of the enzymes will be assembled and screened for specific functional activities. Then variants will be engineered to have improved efficiency -- so that at least 20 percent of the cells targeted will be altered -- and specificity in inserting or deleting genes. The functional domains then will be coupled to signals for recognizing specific genetic sequences for purposes of targeting desired locations within a genome. Finally, a control region that allows researchers to selectively induce enzyme activity will be added. Once developed, the new tools will be demonstrated by excision of mutant genes and insertion of corrected genes in a cancer model, and by gene correction in an animal model for muscular dystrophy. This research could not be undertaken without ATP support because Rheogene is a small company and the technical risk is too high to attract venture capital funding. If successfully developed and commercialized, the new technology will benefit efforts to develop new cell-based therapeutics and drugs as well as advance the fields of functional genomics and tissue engineering. The technology also could make gene knockouts possible in mitochondria, which are linked to metabolic disorders. The improved efficiency is expected to save as much as 10 percent of the estimated $5 billion spent annually on research and development related to gene function and help ensure U.S. leadership in this arena.

For project information:

Lorraine Keller, (215) 619-5526
lkeller@rheogene.com

ATP Project Manager

Mrunal Chapekar, (301) 975-6846
mrunal.chapekar@nist.gov