Cell-free recombinase-integrated Boolean output system

Cell-free gene expression systems are increasingly important in fundamental research and biomanufacturing, offering a versatile platform for studying gene circuits and biocomputation. We present the cell-free recombinase-integrated Boolean output system (CRIBOS), a site-specific recombinase-based multiplex genetic circuit platform designed for cell-free environments. With CRIBOS, we built over 20 multi-input-multi-output circuits, including 2-input-2-output genetic circuits and a 2-input-4-output decoder. Combined with allosteric transcription factor (aTF)-based sensors, the circuits demonstrate multiplex environmental sensing. Moreover, utilizing paper-based CRIBOS, which demonstrates remarkable portability and stability, we present a biological memory storage logic circuit device that can preserve DNA-based biological information for over 4 months with minimal resources, energy costs, and maintenance requirements. Implementing CRIBOS not only expands the application of multiplex Boole..., , # Data from: Cell-free recombinase-integrated Boolean output system Dataset DOI: [10.5061/dryad.xksn02vws](https://doi.org/10.5061/dryad.xksn02vws) ## Description of the data and file structure #### File: Cell_free_paper_Final_Processed_Data_reformatted.xlsx Our datasets, comprised of many recombinased-based genetic circuits designed to operate in a cell-free environment, support the 6 major data figures in our manuscript. Figure 1: Schematic diagrams of cell-free recombinase circuits Figure 2:  Characterization of cell-free recombinase excision circuits Figure 3: Characterization of activation TF-implemented recombinase genetic circuits in a cell-free condition Figure 4: 2-input-2-output circuits for sensing two chemicals in the cell-free condition. The two chemicals are Zn ion and tetracycline (tet). Zn controls the expression of PhiC recombinase, whereas Tet controls the expression of Cre. Figure 5: 2-input-4-output genetic decoder performance in cell free environment ..., ,