Engineering the TetO System to Test the Contribution of FKS1 to Yeast Cell Wall Strength

Abstract

Society has a demand for manufactured proteins such as insulin, which can be produced by the yeast Saccharomyces cerevisiae, but such proteins are not easily accessible due to this yeast’s rigid cell wall. I hypothesize that if S. cerevisiae cells were not able to make as much FKS1 protein (a protein involved in cell wall synthesis) they would exhibit reduced growth rates and weaker cell walls. The approach is to genetically reprogram the yeast to reduce production of FKS1 when exposed to doxycycline. To accomplish this, the native FKS1 and GSC2 genes were knocked out, leaving only the doxycycline regulated FKS1 gene. Compared to the wildtype yeast, the TetO regulated yeast exhibited a reduced growth rate when exposed to doxycycline. In pursuit of heterologous proteins, further experimentation of the TetO system may be considered. Different production platforms may prove more appropriate in future studies due to greater efficacy in reducing cell wall strength.