Cell-free expression systems are becoming robust platforms to reduce system complexity, to use toxic and non-naturally occurring compounds or to produce metabolic intermediates, proteins and molecules that are otherwise toxic to cells. Unfortunately, current cell-free expression platforms are often inconsistent, low scale, not flexible, poorly characterized and limited to relatively simple biological processes, which has prevented their widespread adoption. There is a critical need for a consistent, reliable and commercially available cell-free transcription/translation (TXTL) system that integrates the last progresses made in this area. Here we propose to develop a commercial TXTL offer based on a powerful E. coli based cell-free platform that recently emerged as a promising technology for in vitro prototyping and expression of complex molecular programs. This system is the first to use the endogenous transcription machinery in addition to the T7-based systems, expanding the repertoire of transcription to hundreds of parts. As a consequence, it has a unique versatility for prototyping biomolecular networks. This platform is also highly tunable, both at the transcription and translation level, enabling fine gene circuitry regulations. The commercialization of this technology will enable widespread use of cell-free platforms for rapid prototyping and biomanufacturing.
