CFPU: A Cell-Free Processing Unit for High-Throughput, Automated In Vitro Circuit Characterization in Steady-State Conditions

Forward engineering synthetic circuits are at the core of biology. Automated solutions will be required to facilitate circuit design and implementation. Circuit is increasingly being automated with software, but innovations in experimental automation lagging behind. Microfluidic technologies made it possible perform vitro transcription-translation (tx-tl) reactions increasing throughput sophistication, enabling screening characterization individual elements complete designs. Here, we developed an microfluidic cell-free processing unit (CFPU) that extends high-throughput capabilities a steady-state reaction environment, which essential for implementation analysis more complex dynamic circuits. The CFPU contains 280 chemostats can individually programmed DNA Each chemostat periodically supplied tx-tl reagents, giving rise sustained, long-term conditions. Using pulse width modulation (PWM), device able generate reagent compositions real time. has higher throughput, lower consumption, overall functionality than current devices. We applied this technology map transcription factor-based repression under equilibrium conditions implemented gene switchable by small molecules. expect help bridge gap between development