Meeting Proceedings from the 5th Minimal Cell WorkshopLife at Its Simplest Foundations in Minimal Cells.

The fifth annual virtual minimal cell workshop, hosted by the J. Craig Venter Institute (JCVI), brought together an international community of researchers from academic, industrial, and government laboratories. The 2025 meeting featured 30 talks focused on using minimal bacterial cells to probe the fundamental principles of life, develop new construction strategies, and advance predictive frameworks for cellular behavior. Most of the presentations centered on the JCVI minimal bacterial cells JCVI-syn3.0, JCVI-syn3A, and JCVI-syn3B. Multiple talks addressed the persistent set of essential genes with unknown or partially defined functions, leveraging advanced imaging, computational modeling, and experimental evolution to uncover their cellular roles. Studies of protein moonlighting, membrane biophysics, cell division, metabolism, and aging demonstrated how minimal cells expose fundamental biological constraints often obscured in more complex organisms. Whole-cell computational models increasingly reproduce growth, division, metabolism, gene regulation, and macromolecular assembly. Presentations also highlighted new approaches to whole-genome transplantation, development of alternative minimal genome chassis, synthetic cell assembly from non-living components, chromosome-scale minimization in eukaryotes, and conceptual agricultural and prebiotic systems. Collectively, these efforts position minimal cells as powerful platforms for understanding, building, and predicting living systems.