Ideas Lab: CFIRE: Meta-PURE: End-Use-Driven Cell-Free Modules

Award ID: 2452482

Meta-PURE: End-Use-Driven Cell-Free Modules Cell-free systems reduce biology to its most basic parts, simplifying the complexity of traditional biomanufacturing. However, scientists still do not have an efficient way to plug-and-play these individual cell-free pieces. This project addresses this gap by developing a modular, standardized platform for cell-free bioprocessing. Instead of custom, one-off designs, the project creates eight reusable modules covering essential functions like energy generation and protein expression. These ready-to-use modules make it easier to mix and match capabilities, allowing for faster, more efficient product development. The team will demonstrate how modular design and standardized kits can reduce costs, improve accessibility, and boost productivity across three market-relevant targets. In doing so, this project aims to make cell-free technologies broadly available and expand the use of cell-free technologies across the U.S. bioeconomy. This project will also cultivate the next generation of biotechnology talent through a dynamic, cross-sector team of postdocs and graduate studentsuniting expertise from industry, academia, and governmentto pioneer and scale the future of cell-free biomanufacturing. The project pioneers a modular, standardized framework to address bottlenecks for scaling cell-free bioprocessing. The project will develop, characterize, and integrate eight distinct functional modules common across cell-free systems: three for energy generation, three for the synthesis of valuable products, and two for transcriptiontranslation. These modules span varying levels of complexityfrom individual purified enzymes to enzyme cascades, PUrified Recombinant Elements (PURE), and lysate-based systemsencompassing the full spectrum of cell-free biotechnologies. This modularity allows innovation beyond standalone PURE technologies; it comprises modules with defined capabilities, each built and optimized independently, then integrated to achieve more complex and ambitious synthesis goals. The project also incorporates advanced analytical tools and establishes new standards and metrics to rigorously evaluate module performance and compatibility, which will result in a system that provides reliable synthesis. Further, the system will be validated through the synthesis of three industrially relevant targets: the valuable small molecule santalene; the GamS protein, which enhances cell-free protein expression; and production of a bacteriophage. These demonstrations will show how the system can be used for a varied of use cases. This project will accelerate innovation, enhance reproducibility, and demonstrate a replicable framework that enables flexible, cost-effective, and high-performance bio-based production platforms across diverse applications to support scalable biomanufacturing and strengthen the infrastructure of the U.S. bioeconomy. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NSF Program Director: Waleed Nasser