Innovations in cell-free synthetic biology via microfluidic approaches

Cell-free synthetic biology is a powerful technology that is gaining increasing popularity due to its ability to perform complex biochemical reactions in a well-controlled environment, isolated from the intricacies of living cells. Although it has demonstrated significant success in genetic part characterization and high-throughput protein production without the constraints of cellular membranes, cell-free systems still face several challenges. These include the limited volume available for transcription-translation machinery, the difficulty in standardizing cell lysis procedures, and the variability in cell extract performance across different laboratories. Microfluidic technology has become a powerful tool to address these challenges by supporting the miniaturization and automation of complex, multi-step workflows. Integrating cell-free gene and protein synthesis with microfluidic platforms has redefined bioprocessing, making it more compact and accessible. This synergy has unlocked a wide range of applications across diverse areas of synthetic biology in recent years. This book chapter reviews microfluidic-based methods for synthetic biology in cell-free environments. It highlights the fundamental principles of both cell-free systems and microfluidic technologies, and presents various examples of their integration. The chapter proposes this convergence as a pioneering approach to developing innovative platforms for synthetic biology, with potential applications in biomedical, therapeutic, diagnostic, and environmental contexts.