Initially, we saw little success in our efforts to PCR amplify the DNA components of our gene drive complexes. Eventually, we succeeded in this task, and saw an early success in combining the various components into their respective gene drive complexes. However, this success was not easily reproduced, and much of our effort went into attempting to create the complexes needed. After many trials, it was finally determined that we had nucleases in our water stock which prevented us from amplifying our DNA in order to carry out further cloning steps. We were able to clone one part of our recovery drive, a yeast codon optimized TetR.
After the necessary gene drives were created, our plan was to modify our yeast with both drives using homologous recombination. When that was achieved, the first gene drive would be observed in action. Then, on a population of mutant-type yeast that resulted from the gene drive, the recovery drive would be activated and its effects would be observed.
Future work on this subject could involve experimenting with different promoters for the recovery drive. One early idea was using a light-activated promoter to start the recovery drive. Other potential expansions of this work could involve inserting new genes into the yeast instead of knocking out genes that already exist within the yeast. Through these investigations, more insight could be gained into how gene drives operate and how they can be controlled.