Carefully consider potential leaky expression. Background expression, even without tetracycline, can significantly impact experimental results, especially in sensitive assays. Optimize your system using various tetracycline concentrations to minimize this.
Avoid prolonged inductions. Long-term exposure to tetracycline can lead to decreased induction efficiency and potentially cytotoxic effects. Implement short, controlled inductions whenever possible.
Recognize cell-line specific variations. Induction strength and response kinetics vary between cell lines. Titrate tetracycline concentrations and carefully monitor expression levels for each cell type you use.
Account for potential off-target effects. Tetracycline can influence cellular processes independent of its effect on the promoter. Conduct control experiments to rule out confounding factors.
Address potential dosage effects. High tetracycline concentrations might negatively impact cell viability or cause nonspecific effects. Determine an optimal induction concentration through careful dose-response experiments.
Consider alternative systems. If you encounter significant limitations, explore alternative inducible promoter systems suitable for your specific application. Each system has its own strengths and weaknesses.
Analyze your data critically. Always carefully interpret your results, taking potential limitations of the tetracycline inducible system into account. Statistical analysis and appropriate controls are critical.
