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Our project aims to develop a novel mitochondrial transformation technique utilizing the cotransport of tRNA to import exogenous DNA into the mitochondria of S. cerevisiae cells. Previous work on this project created novel tRNA-DNA hybrid vectors, optimized transformation protocols, and analyzed putative S. cerevisiae mitochondrial transformants. We continued this investigation with the optimization of spheroplast and polymerase chain reaction. We also explored the stability and lifespan of post-transformation yeast cells by analyzing the mitochondrial gene, ATP6. If the cotransport of tRNA-Arg8 is successful, our findings could be applied to human mitochondria, providing a tool to better study and treat diseases.
