Invention:
This technology involves a novel system using a two-step process to eliminate a gene of interest by incorporating a “suicide gene” vector system. The construct created by this method results with no foreign sequences, making the strains markerless mutants. This means that one has the potential to make genetic strains with multiple mutations and can therefore contribute to nearly every area of biological research.
Deletion of the Coccidioides CPS1 gene resulted in a mutant strain that shows efficacy and safety as a liveattenuated vaccine to prevent coccidioidomycosis (Valley Fever) that is being developed for commercial production as a canine vaccine. The markerless cps1 deletion mutant may be critical for getting regulatory approval of a live attenuated cps1 vaccine for humans.
Background:
Ultimately, one wishes to determine how genes—and the proteins they encode—function in the intact organism. Although it may sound counterintuitive, one of the most direct ways to find out what a gene does is to see what happens to the organism when that gene is missing. Studying mutant organisms that have acquired changes or deletions in their nucleotide sequences is a time-honored practice in biology. Because mutations can interrupt cellular processes, mutants often hold the key to understanding gene function. In the classical approach to the important field of genetics, one begins by isolating mutants that have an interesting or unusual appearance: fruit flies with white eyes or curly wings, for example. While methods of producing these mutations have become more sophisticated as time has progressed, the need to modernize biological techniques has never been greater.
Applications:
- Biological research
- Vaccine research using the CPS1 deletion mutant
- Potential vaccine for coccidioidomycosis (Valley Fever)
Advantages:
- Researchers can create multi-mutated strains
- No foreign DNA is introduced into the genetic constructs formed by using this method
