Temperature-Dependent Recombinase-Based Genetic Circuits

Temperature offers a simple yet powerful signal to program cellular behavior. Here, we engineered and characterized a set of temperature-dependent genetic circuits that integrate RNA thermometers with site-specific DNA recombinases to achieve precise, irreversible control of gene expression. Using the serine recombinase Bxb1 placed under the control of the Salmonella FourU RNA thermometer, we demonstrate how promoter strength critically defines thermal sensitivity: weak promoters activity clear ON/OFF transitions, while strong promoters lead to continuous, quasi temperature-independent recombination. Furthermore, temperature pulse duration and growth phase of cell culture were found to modulate recombination efficiency, providing additional layers of control. We illustrate the potential of this framework through proof-of-concept applications, including (i) the generation of spatial expression patterns on 2D surfaces via localized heating, (ii) a paper-based device capable of recording temperature gradients as stable genetic outputs, and (iii) a temperature-triggered lysis system for controlled cellular release. Together, these results establish temperature-regulated recombinase circuits as versatile and robust tools for programmable, spatially resolved, and irreversible control of gene expression, paving the way for new applications in synthetic biology, biosensing, and bioproduction.