Methods and Applications for Controlling Cellular Viability Using Fluoride-Sensitivity
Tech ID: 31838 / UC Case 2018-424-0
This technology employs fluoride-sensitivity to overcome the limitations of existing selection methods.
Molecular and genetic technologies require the ability to select for specific cellular functions. Antibiotic and auxotrophic selection methods are commonly used in protein and metabolic engineering and synthetic biology, but they pose significant limitations. Antibiotic selection generates antibiotic-resistant microorganisms and the antibiotics themselves can be expensive. Auxotrophic selection requires the use of defined media, where some organisms will become nutrient-limited as defined media is generally less rich in nutrients than complex media. A selection method that overcomes the expense and media limitations of current methods would be widely applicable in protein and metabolic engineering and synthetic biology applications.
Researchers at the University of California, Santa Barbara have employed fluoride-sensitivity to screen for and introduce desired cellular properties. This process does not require defined media or expensive antibiotics and there is no possibility of antibiotic resistance. Fluoride acts as a toxic molecule to many organisms, and the defensive measures that cells take to protect themselves from environmental fluoride are foundational for screening. Removing or adding the mechanism that protects cells from fluoride — fluoride transport (FEX) proteins — enables selection of desirable properties. In addition to serving as a selection platform, fluoride-sensitivity enables a stringent, easily implemented, and versatile biocontainment strategy. This system addresses the scarcity of platforms available for yeast despite their prevalent use in industry and academic research.
- Tunable selection pressure
- No costly antibiotics or chance of antibiotic resistance
- Allows for complex media
- Metabolic engineering
- Protein engineering
- Synthetic biology
- Protein production